In this notebook resides all of my code for analizing our sequences in terms of their ARE score distributions... the first code block will append our ARE score information and boolian tissue presence indicators for use in upset plotting and ARE expression differential analysis.

In [3]:
import pandas as pd
import os

# Function to read sequences from a FASTA file into a dictionary
def read_fasta(file_path):
    sequences = {}
    with open(file_path, 'r') as f:
        gene_id = ""
        transcript_id = ""
        sequence = ""
        for line in f:
            if line.startswith(">"):
                if gene_id and sequence:
                    if gene_id not in sequences:
                        sequences[gene_id] = {}
                    sequences[gene_id][transcript_id] = sequence
                header_parts = line.strip().lstrip(">").split('|')
                gene_id = header_parts[0]
                transcript_id = header_parts[1]
                sequence = ""
            else:
                sequence += line.strip()
        if gene_id and sequence:
            if gene_id not in sequences:
                sequences[gene_id] = {}
            sequences[gene_id][transcript_id] = sequence
    return sequences

def parse_are_score(file_path, longest_transcripts, sequences):
    print(f"Parsing ARE score data from {file_path}...")
    with open(file_path, 'r') as f:
        lines = f.readlines()[1:]  # Skip header
        for line in lines:
            parts = line.strip().split('\t')
            gene_id = parts[0].split('|')[0]
            transcript_id = parts[0].split('|')[1]
            score = float(parts[1])
            pentamer_count = int(parts[2])
            pentamer_position = parts[3]
            au_block_position = parts[4]
            sequence_length = int(parts[5])

            sequence = ""
            if gene_id in sequences and transcript_id in sequences[gene_id]:
                sequence = sequences[gene_id][transcript_id]
            else:
                print(f"Warning: No sequence found for {transcript_id}")

            # Check if we should update the longest transcript information for this gene
            if gene_id not in longest_transcripts or sequence_length > longest_transcripts[gene_id]['Sequence_length']:
                longest_transcripts[gene_id] = {
                    'ARE_score': score,
                    'Pentamer_count': pentamer_count,
                    'Pentamer_position': pentamer_position,
                    'AU_block_position': au_block_position,
                    'Sequence': sequence,
                    'Sequence_length': sequence_length
                }
                print(f"Updated longest transcript for {gene_id}: {sequence_length}")
    print(f"Completed parsing {file_path}. Total genes: {len(longest_transcripts)}")
    return longest_transcripts

def count_motifs(lengths):
    motif_7mer = "TATTTAT"
    motif_5mer = "ATTTA"
    motif_counts = {'7mer_count': {}, '5mer_count': {}}
    for key, val in lengths.items():
        i_count_7mer, i_count_5mer = 0, 0
        sequence = val['Sequence']

        for i in range(0, len(sequence) - len(motif_7mer) + 1):
            if sequence[i:i + len(motif_7mer)] == motif_7mer:
                i_count_7mer += 1
        for i in range(0, len(sequence) - len(motif_5mer) + 1):
            if sequence[i:i + len(motif_5mer)] == motif_5mer:
                i_count_5mer += 1
        motif_counts['7mer_count'][key] = i_count_7mer
        motif_counts['5mer_count'][key] = i_count_5mer
    return motif_counts

def update_annotated_files(input_dir, output_dir, regulation, are_score_data, motif_counts):
    if not os.path.exists(output_dir):
        os.makedirs(output_dir)
    files = [f for f in os.listdir(input_dir) if f.endswith("_annotated.csv")]
    total_files = len(files)
    print(f"Updating {total_files} `{regulation}` files in {input_dir}...")
    gene_presence = {tissue: set() for tissue in ["BM", "Spleen", "Ovary", "Liver", "Lung", "Kidney"]}
    # First pass: identify the presence of each gene in each tissue
    for file in files:
        input_file_path = os.path.join(input_dir, file)
        df = pd.read_csv(input_file_path)
        tissue = file.split('_')[0]
        for gene_id in df['ID']:
            gene_presence[tissue].add(gene_id)
    # Update the files with ARE score data and Boolean indicators
    for idx, file in enumerate(files):
        input_file_path = os.path.join(input_dir, file)
        df = pd.read_csv(input_file_path)
        # Extract tissue type from filename
        tissue = file.split('_')[0]
        # Create new columns
        df['ARE_score'] = df['ID'].map(lambda x: are_score_data.get(x, {}).get('ARE_score', 'N/A'))
        df['Pentamer_count'] = df['ID'].map(lambda x: are_score_data.get(x, {}).get('Pentamer_count', 'N/A'))
        df['Pentamer_position'] = df['ID'].map(lambda x: are_score_data.get(x, {}).get('Pentamer_position', 'N/A'))
        df['AU_block_position'] = df['ID'].map(lambda x: are_score_data.get(x, {}).get('AU_block_position', 'N/A'))
        df['Sequence_length'] = df['ID'].map(lambda x: are_score_data.get(x, {}).get('Sequence_length', 'N/A'))
        df['5mer_count'] = df['ID'].map(lambda x: motif_counts['5mer_count'].get(x, 'N/A'))
        df['7mer_count'] = df['ID'].map(lambda x: motif_counts['7mer_count'].get(x, 'N/A'))
        # Add Boolean indicators for each tissue
        df['BM'] = df['ID'].map(lambda x: 1 if x in gene_presence['BM'] else 0)
        df['Spleen'] = df['ID'].map(lambda x: 1 if x in gene_presence['Spleen'] else 0)
        df['Ovary'] = df['ID'].map(lambda x: 1 if x in gene_presence['Ovary'] else 0)
        df['Liver'] = df['ID'].map(lambda x: 1 if x in gene_presence['Liver'] else 0)
        df['Lung'] = df['ID'].map(lambda x: 1 if x in gene_presence['Lung'] else 0)
        df['Kidney'] = df['ID'].map(lambda x: 1 if x in gene_presence['Kidney'] else 0)
        # Save the updated file
        output_file_path = os.path.join(output_dir, file.replace("_annotated.csv", "_annotated_updated.csv"))
        df.to_csv(output_file_path, index=False)
        # Print progress information
        print(f"Processed {idx + 1}/{total_files} files. Current file: {file}")
    print(f"Completed updating files in {input_dir}.")

# Read sequences from the FASTA files
print("Reading sequences from FASTA files...")
sequences_38 = read_fasta('/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE/sequences/GRC_38p6_full.fasta')
# sequences_39 = read_fasta('/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE/sequences/GRC39_full.fasta')
# Combine sequences dictionaries, assuming unique transcript IDs across the versions
all_sequences = {**sequences_38}

# Initialize the dictionary for ARE score data
are_score_data = {}
# Parse ARE score data from both the GRC38 and GRC39 files
print("Parsing ARE score data from txt files...")
are_score_data = parse_are_score('ARE_score_data/3primeUTR_AREscore_GRC38p6_full.txt', are_score_data, all_sequences)
are_score_data = parse_are_score('ARE_score_data/3primeUTR_AREscore_GRC39_full.txt', are_score_data, all_sequences)

# Calculate motif counts
motif_counts = count_motifs(are_score_data)

print("Completed parsing ARE score data and calculating motif counts.")
# Update the upregulated and downregulated files
update_annotated_files("upregulated_genes", "upregulated_genes_updated", "upregulated", are_score_data, motif_counts)
update_annotated_files("unregulated_genes", "unregulated_genes_updated", "unregulated", are_score_data, motif_counts)
update_annotated_files("downregulated_genes", "downregulated_genes_updated", "downregulated", are_score_data, motif_counts)
print("All files have been updated successfully.")
Reading sequences from FASTA files...
Parsing ARE score data from txt files...
Parsing ARE score data from ARE_score_data/3primeUTR_AREscore_GRC38p6_full.txt...
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Updated longest transcript for ENSMUSG00000033510: 297
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Updated longest transcript for ENSMUSG00000116094: 533
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Updated longest transcript for ENSMUSG00000027134: 224
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Updated longest transcript for ENSMUSG00000039865: 444
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Updated longest transcript for ENSMUSG00000026511: 464
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Updated longest transcript for ENSMUSG00000079389: 319
Updated longest transcript for ENSMUSG00000070390: 7
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Updated longest transcript for ENSMUSG00000027510: 668
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Updated longest transcript for ENSMUSG00000060450: 266
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Updated longest transcript for ENSMUSG00000042607: 1975
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Updated longest transcript for ENSMUSG00000045404: 789
Updated longest transcript for ENSMUSG00000028431: 1972
Updated longest transcript for ENSMUSG00000028850: 411
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Updated longest transcript for ENSMUSG00000024403: 969
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Updated longest transcript for ENSMUSG00000074480: 4215
Updated longest transcript for ENSMUSG00000052713: 3720
Updated longest transcript for ENSMUSG00000073257: 103
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Updated longest transcript for ENSMUSG00000097395: 366
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Updated longest transcript for ENSMUSG00000020863: 1979
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Updated longest transcript for ENSMUSG00000089960: 573
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Updated longest transcript for ENSMUSG00000029598: 2389
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Updated longest transcript for ENSMUSG00000039904: 1538
Updated longest transcript for ENSMUSG00000021947: 456
Updated longest transcript for ENSMUSG00000054408: 2830
Updated longest transcript for ENSMUSG00000021832: 1004
Updated longest transcript for ENSMUSG00000032399: 222
Updated longest transcript for ENSMUSG00000074892: 3764
Updated longest transcript for ENSMUSG00000050996: 669
Updated longest transcript for ENSMUSG00000034401: 818
Updated longest transcript for ENSMUSG00000024300: 1333
Updated longest transcript for ENSMUSG00000070565: 5529
Updated longest transcript for ENSMUSG00000056536: 3582
Updated longest transcript for ENSMUSG00000002635: 182
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Updated longest transcript for ENSMUSG00000001506: 1423
Updated longest transcript for ENSMUSG00000030584: 1037
Updated longest transcript for ENSMUSG00000047420: 857
Updated longest transcript for ENSMUSG00000035762: 1710
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Updated longest transcript for ENSMUSG00000013150: 209
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Updated longest transcript for ENSMUSG00000024331: 3121
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Updated longest transcript for ENSMUSG00000026956: 1758
Updated longest transcript for ENSMUSG00000111842: 161
Updated longest transcript for ENSMUSG00000045474: 229
Updated longest transcript for ENSMUSG00000071065: 2403
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Updated longest transcript for ENSMUSG00000064145: 2140
Updated longest transcript for ENSMUSG00000015083: 159
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Updated longest transcript for ENSMUSG00000039512: 492
Updated longest transcript for ENSMUSG00000039640: 631
Updated longest transcript for ENSMUSG00000035403: 540
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Updated longest transcript for ENSMUSG00000052833: 787
Updated longest transcript for ENSMUSG00000008590: 892
Updated longest transcript for ENSMUSG00000021884: 1182
Updated longest transcript for ENSMUSG00000060201: 219
Updated longest transcript for ENSMUSG00000090363: 1188
Updated longest transcript for ENSMUSG00000026975: 548
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Updated longest transcript for ENSMUSG00000052435: 179
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Updated longest transcript for ENSMUSG00000040564: 113
Updated longest transcript for ENSMUSG00000073557: 573
Updated longest transcript for ENSMUSG00000035946: 587
Updated longest transcript for ENSMUSG00000025789: 4060
Updated longest transcript for ENSMUSG00000038594: 42
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Updated longest transcript for ENSMUSG00000095887: 105
Updated longest transcript for ENSMUSG00000029730: 136
Updated longest transcript for ENSMUSG00000019843: 1409
Updated longest transcript for ENSMUSG00000007783: 127
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Updated longest transcript for ENSMUSG00000028114: 2910
Updated longest transcript for ENSMUSG00000095518: 761
Updated longest transcript for ENSMUSG00000032555: 512
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Updated longest transcript for ENSMUSG00000079362: 2398
Updated longest transcript for ENSMUSG00000038175: 1512
Updated longest transcript for ENSMUSG00000021408: 4365
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Updated longest transcript for ENSMUSG00000029419: 1557
Updated longest transcript for ENSMUSG00000068732: 2453
Updated longest transcript for ENSMUSG00000027751: 1982
Updated longest transcript for ENSMUSG00000028382: 5015
Updated longest transcript for ENSMUSG00000067591: 315
Updated longest transcript for ENSMUSG00000030753: 1159
Updated longest transcript for ENSMUSG00000023073: 2937
Updated longest transcript for ENSMUSG00000031826: 790
Updated longest transcript for ENSMUSG00000050788: 1470
Updated longest transcript for ENSMUSG00000038132: 1778
Updated longest transcript for ENSMUSG00000028329: 310
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Updated longest transcript for ENSMUSG00000052276: 817
Updated longest transcript for ENSMUSG00000057378: 381
Updated longest transcript for ENSMUSG00000078350: 3124
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Updated longest transcript for ENSMUSG00000024446: 174
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Updated longest transcript for ENSMUSG00000013846: 372
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Updated longest transcript for ENSMUSG00000038085: 188
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Updated longest transcript for ENSMUSG00000068037: 481
Updated longest transcript for ENSMUSG00000111709: 105
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Updated longest transcript for ENSMUSG00000002297: 239
Updated longest transcript for ENSMUSG00000096273: 241
Updated longest transcript for ENSMUSG00000022790: 1964
Updated longest transcript for ENSMUSG00000033420: 4309
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Updated longest transcript for ENSMUSG00000059495: 5340
Updated longest transcript for ENSMUSG00000010914: 1402
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Updated longest transcript for ENSMUSG00000033916: 213
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Updated longest transcript for ENSMUSG00000098377: 1001
Updated longest transcript for ENSMUSG00000058192: 10938
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Updated longest transcript for ENSMUSG00000022240: 389
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Updated longest transcript for ENSMUSG00000001119: 767
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Updated longest transcript for ENSMUSG00000033282: 4383
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Updated longest transcript for ENSMUSG00000046845: 51
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Updated longest transcript for ENSMUSG00000032280: 1781
Updated longest transcript for ENSMUSG00000049152: 612
Updated longest transcript for ENSMUSG00000060572: 404
Updated longest transcript for ENSMUSG00000053846: 1995
Updated longest transcript for ENSMUSG00000033972: 797
Updated longest transcript for ENSMUSG00000116418: 99
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Updated longest transcript for ENSMUSG00000079444: 989
Updated longest transcript for ENSMUSG00000066896: 1771
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Updated longest transcript for ENSMUSG00000039196: 113
Updated longest transcript for ENSMUSG00000055471: 470
Updated longest transcript for ENSMUSG00000096629: 1133
Updated longest transcript for ENSMUSG00000027208: 1624
Updated longest transcript for ENSMUSG00000015659: 1918
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Updated longest transcript for ENSMUSG00000032641: 186
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Updated longest transcript for ENSMUSG00000019782: 172
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Updated longest transcript for ENSMUSG00000041959: 239
Updated longest transcript for ENSMUSG00000025417: 1997
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Updated longest transcript for ENSMUSG00000008482: 397
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Updated longest transcript for ENSMUSG00000001687: 1500
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Updated longest transcript for ENSMUSG00000043982: 26
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Updated longest transcript for ENSMUSG00000041138: 239
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Updated longest transcript for ENSMUSG00000013150: 2898
Updated longest transcript for ENSMUSG00000091955: 285
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Updated longest transcript for ENSMUSG00000013846: 3782
Updated longest transcript for ENSMUSG00000096596: 393
Updated longest transcript for ENSMUSG00000058093: 1568
Updated longest transcript for ENSMUSG00000073755: 1998
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Updated longest transcript for ENSMUSG00000000560: 4205
Updated longest transcript for ENSMUSG00000041328: 1200
Updated longest transcript for ENSMUSG00000059273: 2186
Updated longest transcript for ENSMUSG00000020412: 2621
Updated longest transcript for ENSMUSG00000074698: 2768
Updated longest transcript for ENSMUSG00000033834: 1114
Updated longest transcript for ENSMUSG00000020945: 252
Updated longest transcript for ENSMUSG00000056617: 330
Updated longest transcript for ENSMUSG00000030345: 640
Updated longest transcript for ENSMUSG00000032911: 980
Updated longest transcript for ENSMUSG00000037656: 1192
Updated longest transcript for ENSMUSG00000024912: 638
Updated longest transcript for ENSMUSG00000052456: 467
Updated longest transcript for ENSMUSG00000026526: 1050
Updated longest transcript for ENSMUSG00000000378: 383
Updated longest transcript for ENSMUSG00000066151: 1750
Updated longest transcript for ENSMUSG00000004631: 757
Updated longest transcript for ENSMUSG00000046959: 1339
Updated longest transcript for ENSMUSG00000044906: 1620
Updated longest transcript for ENSMUSG00000027438: 3913
Updated longest transcript for ENSMUSG00000047139: 1500
Updated longest transcript for ENSMUSG00000029247: 695
Updated longest transcript for ENSMUSG00000042258: 1199
Updated longest transcript for ENSMUSG00000020168: 3152
Updated longest transcript for ENSMUSG00000050555: 354
Updated longest transcript for ENSMUSG00000067399: 345
Updated longest transcript for ENSMUSG00000026546: 117
Updated longest transcript for ENSMUSG00000074849: 797
Updated longest transcript for ENSMUSG00000095015: 1366
Updated longest transcript for ENSMUSG00000067704: 716
Updated longest transcript for ENSMUSG00000095609: 154
Updated longest transcript for ENSMUSG00000040767: 165
Updated longest transcript for ENSMUSG00000024983: 3399
Updated longest transcript for ENSMUSG00000036557: 25
Updated longest transcript for ENSMUSG00000024187: 1684
Updated longest transcript for ENSMUSG00000079169: 44
Updated longest transcript for ENSMUSG00000096488: 1075
Updated longest transcript for ENSMUSG00000071037: 449
Updated longest transcript for ENSMUSG00000027669: 4788
Updated longest transcript for ENSMUSG00000075272: 432
Updated longest transcript for ENSMUSG00000095474: 38
Updated longest transcript for ENSMUSG00000039206: 2543
Updated longest transcript for ENSMUSG00000035944: 2258
Updated longest transcript for ENSMUSG00000003575: 3769
Updated longest transcript for ENSMUSG00000029586: 909
Updated longest transcript for ENSMUSG00000023078: 800
Updated longest transcript for ENSMUSG00000022360: 1481
Updated longest transcript for ENSMUSG00000058398: 821
Updated longest transcript for ENSMUSG00000074366: 803
Updated longest transcript for ENSMUSG00000095687: 1585
Updated longest transcript for ENSMUSG00000094496: 1060
Updated longest transcript for ENSMUSG00000033004: 12315
Updated longest transcript for ENSMUSG00000103707: 3817
Updated longest transcript for ENSMUSG00000032299: 414
Updated longest transcript for ENSMUSG00000027406: 331
Updated longest transcript for ENSMUSG00000028542: 2221
Updated longest transcript for ENSMUSG00000050224: 261
Updated longest transcript for ENSMUSG00000033813: 1545
Updated longest transcript for ENSMUSG00000116336: 759
Updated longest transcript for ENSMUSG00000041189: 585
Updated longest transcript for ENSMUSG00000078173: 1466
Updated longest transcript for ENSMUSG00000096641: 296
Updated longest transcript for ENSMUSG00000004054: 1254
Updated longest transcript for ENSMUSG00000052504: 2597
Updated longest transcript for ENSMUSG00000029390: 1301
Updated longest transcript for ENSMUSG00000000093: 934
Updated longest transcript for ENSMUSG00000072568: 3969
Updated longest transcript for ENSMUSG00000021866: 734
Updated longest transcript for ENSMUSG00000105504: 1515
Updated longest transcript for ENSMUSG00000041079: 985
Updated longest transcript for ENSMUSG00000038725: 199
Updated longest transcript for ENSMUSG00000023039: 2648
Updated longest transcript for ENSMUSG00000087247: 474
Updated longest transcript for ENSMUSG00000020211: 110
Updated longest transcript for ENSMUSG00000026834: 7465
Updated longest transcript for ENSMUSG00000044595: 360
Updated longest transcript for ENSMUSG00000006488: 625
Updated longest transcript for ENSMUSG00000027803: 3277
Updated longest transcript for ENSMUSG00000051728: 1878
Updated longest transcript for ENSMUSG00000021056: 107
Updated longest transcript for ENSMUSG00000035686: 597
Updated longest transcript for ENSMUSG00000026674: 5548
Updated longest transcript for ENSMUSG00000095028: 208
Updated longest transcript for ENSMUSG00000042115: 2565
Updated longest transcript for ENSMUSG00000026645: 428
Updated longest transcript for ENSMUSG00000094152: 321
Updated longest transcript for ENSMUSG00000060962: 324
Updated longest transcript for ENSMUSG00000031711: 608
Updated longest transcript for ENSMUSG00000091476: 2894
Updated longest transcript for ENSMUSG00000081534: 1969
Updated longest transcript for ENSMUSG00000020191: 92
Updated longest transcript for ENSMUSG00000048677: 1273
Updated longest transcript for ENSMUSG00000027533: 487
Updated longest transcript for ENSMUSG00000058145: 3105
Updated longest transcript for ENSMUSG00000053279: 499
Updated longest transcript for ENSMUSG00000040629: 347
Updated longest transcript for ENSMUSG00000035277: 954
Updated longest transcript for ENSMUSG00000006345: 309
Updated longest transcript for ENSMUSG00000016252: 153
Updated longest transcript for ENSMUSG00000020325: 1155
Updated longest transcript for ENSMUSG00000060499: 334
Updated longest transcript for ENSMUSG00000090304: 7343
Updated longest transcript for ENSMUSG00000056234: 3619
Updated longest transcript for ENSMUSG00000034274: 129
Updated longest transcript for ENSMUSG00000051998: 99
Updated longest transcript for ENSMUSG00000045975: 4147
Updated longest transcript for ENSMUSG00000023087: 1112
Updated longest transcript for ENSMUSG00000030742: 225
Updated longest transcript for ENSMUSG00000097328: 590
Updated longest transcript for ENSMUSG00000032549: 4030
Updated longest transcript for ENSMUSG00000036196: 398
Updated longest transcript for ENSMUSG00000030747: 888
Updated longest transcript for ENSMUSG00000045575: 2579
Updated longest transcript for ENSMUSG00000083628: 327
Updated longest transcript for ENSMUSG00000073555: 1205
Updated longest transcript for ENSMUSG00000025226: 155
Updated longest transcript for ENSMUSG00000003882: 2569
Updated longest transcript for ENSMUSG00000035696: 3384
Updated longest transcript for ENSMUSG00000053046: 2355
Updated longest transcript for ENSMUSG00000046474: 309
Updated longest transcript for ENSMUSG00000027456: 418
Updated longest transcript for ENSMUSG00000091712: 3521
Updated longest transcript for ENSMUSG00000027864: 3124
Updated longest transcript for ENSMUSG00000038742: 74
Updated longest transcript for ENSMUSG00000028519: 3347
Updated longest transcript for ENSMUSG00000097351: 1015
Updated longest transcript for ENSMUSG00000029482: 1057
Updated longest transcript for ENSMUSG00000001225: 1813
Updated longest transcript for ENSMUSG00000030368: 17
Updated longest transcript for ENSMUSG00000036231: 190
Updated longest transcript for ENSMUSG00000028525: 1920
Updated longest transcript for ENSMUSG00000037933: 3667
Updated longest transcript for ENSMUSG00000056972: 436
Updated longest transcript for ENSMUSG00000024925: 589
Updated longest transcript for ENSMUSG00000028218: 486
Updated longest transcript for ENSMUSG00000026322: 3282
Updated longest transcript for ENSMUSG00000028455: 475
Updated longest transcript for ENSMUSG00000022229: 629
Updated longest transcript for ENSMUSG00000037971: 1028
Updated longest transcript for ENSMUSG00000028931: 2293
Updated longest transcript for ENSMUSG00000026987: 1286
Updated longest transcript for ENSMUSG00000074832: 1136
Updated longest transcript for ENSMUSG00000040767: 242
Updated longest transcript for ENSMUSG00000042182: 1307
Updated longest transcript for ENSMUSG00000026887: 1495
Updated longest transcript for ENSMUSG00000110344: 1654
Updated longest transcript for ENSMUSG00000040268: 2524
Updated longest transcript for ENSMUSG00000070891: 993
Updated longest transcript for ENSMUSG00000079260: 1298
Updated longest transcript for ENSMUSG00000025577: 2100
Updated longest transcript for ENSMUSG00000037816: 2172
Updated longest transcript for ENSMUSG00000035293: 4264
Updated longest transcript for ENSMUSG00000044279: 622
Updated longest transcript for ENSMUSG00000047819: 1713
Updated longest transcript for ENSMUSG00000021337: 400
Updated longest transcript for ENSMUSG00000055945: 2176
Updated longest transcript for ENSMUSG00000015396: 1447
Updated longest transcript for ENSMUSG00000022234: 128
Updated longest transcript for ENSMUSG00000045613: 3804
Updated longest transcript for ENSMUSG00000019763: 1554
Updated longest transcript for ENSMUSG00000045917: 2392
Updated longest transcript for ENSMUSG00000038080: 2321
Updated longest transcript for ENSMUSG00000053101: 2004
Updated longest transcript for ENSMUSG00000030982: 1672
Updated longest transcript for ENSMUSG00000000782: 1512
Updated longest transcript for ENSMUSG00000026828: 926
Updated longest transcript for ENSMUSG00000055523: 585
Updated longest transcript for ENSMUSG00000075133: 1982
Updated longest transcript for ENSMUSG00000028492: 173
Updated longest transcript for ENSMUSG00000027099: 1957
Updated longest transcript for ENSMUSG00000029623: 1548
Updated longest transcript for ENSMUSG00000030510: 1849
Updated longest transcript for ENSMUSG00000022101: 527
Updated longest transcript for ENSMUSG00000074403: 37
Updated longest transcript for ENSMUSG00000004994: 686
Updated longest transcript for ENSMUSG00000044288: 3796
Updated longest transcript for ENSMUSG00000036216: 232
Updated longest transcript for ENSMUSG00000021279: 1125
Updated longest transcript for ENSMUSG00000004709: 2434
Updated longest transcript for ENSMUSG00000026068: 2617
Updated longest transcript for ENSMUSG00000042492: 1112
Updated longest transcript for ENSMUSG00000045503: 3741
Updated longest transcript for ENSMUSG00000043274: 4190
Updated longest transcript for ENSMUSG00000028540: 1772
Updated longest transcript for ENSMUSG00000027858: 3350
Updated longest transcript for ENSMUSG00000055172: 463
Updated longest transcript for ENSMUSG00000093962: 122
Updated longest transcript for ENSMUSG00000020299: 1434
Updated longest transcript for ENSMUSG00000040731: 1788
Updated longest transcript for ENSMUSG00000050288: 1614
Updated longest transcript for ENSMUSG00000030733: 1520
Updated longest transcript for ENSMUSG00000028187: 1324
Updated longest transcript for ENSMUSG00000051984: 352
Updated longest transcript for ENSMUSG00000040605: 6041
Updated longest transcript for ENSMUSG00000075149: 640
Updated longest transcript for ENSMUSG00000025512: 2912
Updated longest transcript for ENSMUSG00000052298: 2285
Updated longest transcript for ENSMUSG00000021494: 185
Updated longest transcript for ENSMUSG00000038255: 1770
Updated longest transcript for ENSMUSG00000042246: 2296
Updated longest transcript for ENSMUSG00000090486: 780
Updated longest transcript for ENSMUSG00000056608: 3581
Updated longest transcript for ENSMUSG00000024440: 1716
Updated longest transcript for ENSMUSG00000034771: 309
Updated longest transcript for ENSMUSG00000040260: 158
Updated longest transcript for ENSMUSG00000041134: 4657
Updated longest transcript for ENSMUSG00000026222: 2798
Updated longest transcript for ENSMUSG00000096045: 2120
Updated longest transcript for ENSMUSG00000006576: 3282
Updated longest transcript for ENSMUSG00000022297: 1634
Updated longest transcript for ENSMUSG00000068479: 2754
Updated longest transcript for ENSMUSG00000069793: 2786
Updated longest transcript for ENSMUSG00000099564: 435
Updated longest transcript for ENSMUSG00000045795: 760
Updated longest transcript for ENSMUSG00000033027: 364
Updated longest transcript for ENSMUSG00000060510: 3997
Updated longest transcript for ENSMUSG00000045204: 1610
Updated longest transcript for ENSMUSG00000078865: 203
Updated longest transcript for ENSMUSG00000094484: 2026
Updated longest transcript for ENSMUSG00000049551: 313
Updated longest transcript for ENSMUSG00000037509: 1480
Updated longest transcript for ENSMUSG00000032417: 248
Updated longest transcript for ENSMUSG00000028841: 274
Updated longest transcript for ENSMUSG00000070547: 2997
Updated longest transcript for ENSMUSG00000038812: 298
Updated longest transcript for ENSMUSG00000029169: 521
Updated longest transcript for ENSMUSG00000056313: 1396
Updated longest transcript for ENSMUSG00000109129: 330
Updated longest transcript for ENSMUSG00000046192: 1192
Updated longest transcript for ENSMUSG00000042485: 580
Updated longest transcript for ENSMUSG00000020437: 2670
Updated longest transcript for ENSMUSG00000093968: 763
Updated longest transcript for ENSMUSG00000040576: 268
Updated longest transcript for ENSMUSG00000004947: 1240
Updated longest transcript for ENSMUSG00000038541: 751
Updated longest transcript for ENSMUSG00000061778: 959
Updated longest transcript for ENSMUSG00000078799: 110
Updated longest transcript for ENSMUSG00000090942: 1919
Updated longest transcript for ENSMUSG00000001134: 308
Updated longest transcript for ENSMUSG00000032405: 3600
Updated longest transcript for ENSMUSG00000091311: 170
Updated longest transcript for ENSMUSG00000061048: 1847
Updated longest transcript for ENSMUSG00000022911: 1947
Updated longest transcript for ENSMUSG00000094825: 1133
Updated longest transcript for ENSMUSG00000032411: 5675
Updated longest transcript for ENSMUSG00000054434: 992
Updated longest transcript for ENSMUSG00000110591: 1292
Updated longest transcript for ENSMUSG00000024587: 2077
Updated longest transcript for ENSMUSG00000075151: 67
Updated longest transcript for ENSMUSG00000038257: 4718
Updated longest transcript for ENSMUSG00000026100: 1782
Updated longest transcript for ENSMUSG00000031532: 724
Updated longest transcript for ENSMUSG00000073664: 7336
Updated longest transcript for ENSMUSG00000020657: 3672
Updated longest transcript for ENSMUSG00000029070: 758
Updated longest transcript for ENSMUSG00000026102: 4241
Updated longest transcript for ENSMUSG00000029231: 3115
Updated longest transcript for ENSMUSG00000020609: 938
Updated longest transcript for ENSMUSG00000001168: 560
Updated longest transcript for ENSMUSG00000034584: 11
Updated longest transcript for ENSMUSG00000031378: 1003
Updated longest transcript for ENSMUSG00000020072: 1060
Updated longest transcript for ENSMUSG00000030589: 2459
Updated longest transcript for ENSMUSG00000060560: 589
Updated longest transcript for ENSMUSG00000038540: 2667
Updated longest transcript for ENSMUSG00000031073: 1005
Updated longest transcript for ENSMUSG00000051998: 687
Updated longest transcript for ENSMUSG00000027808: 4011
Updated longest transcript for ENSMUSG00000020333: 3559
Updated longest transcript for ENSMUSG00000015023: 1033
Updated longest transcript for ENSMUSG00000058952: 194
Updated longest transcript for ENSMUSG00000022113: 1661
Updated longest transcript for ENSMUSG00000053870: 1698
Updated longest transcript for ENSMUSG00000095075: 635
Updated longest transcript for ENSMUSG00000017049: 174
Updated longest transcript for ENSMUSG00000052752: 2298
Updated longest transcript for ENSMUSG00000094893: 522
Updated longest transcript for ENSMUSG00000000214: 850
Updated longest transcript for ENSMUSG00000062867: 65
Updated longest transcript for ENSMUSG00000038246: 76
Updated longest transcript for ENSMUSG00000036916: 1676
Updated longest transcript for ENSMUSG00000033831: 2262
Updated longest transcript for ENSMUSG00000026343: 978
Updated longest transcript for ENSMUSG00000061845: 88
Updated longest transcript for ENSMUSG00000008958: 273
Updated longest transcript for ENSMUSG00000023176: 2860
Updated longest transcript for ENSMUSG00000052727: 4360
Updated longest transcript for ENSMUSG00000035067: 3686
Updated longest transcript for ENSMUSG00000028702: 245
Updated longest transcript for ENSMUSG00000033080: 2299
Updated longest transcript for ENSMUSG00000079396: 1318
Updated longest transcript for ENSMUSG00000060121: 694
Updated longest transcript for ENSMUSG00000032402: 3495
Updated longest transcript for ENSMUSG00000031661: 2732
Updated longest transcript for ENSMUSG00000029433: 2649
Updated longest transcript for ENSMUSG00000049972: 340
Updated longest transcript for ENSMUSG00000021048: 227
Updated longest transcript for ENSMUSG00000047822: 260
Updated longest transcript for ENSMUSG00000073598: 2021
Updated longest transcript for ENSMUSG00000000204: 922
Updated longest transcript for ENSMUSG00000008348: 92
Updated longest transcript for ENSMUSG00000006398: 178
Updated longest transcript for ENSMUSG00000029408: 1133
Updated longest transcript for ENSMUSG00000026313: 4568
Updated longest transcript for ENSMUSG00000059406: 152
Updated longest transcript for ENSMUSG00000031608: 2286
Updated longest transcript for ENSMUSG00000034472: 1865
Updated longest transcript for ENSMUSG00000026721: 5042
Updated longest transcript for ENSMUSG00000045624: 729
Updated longest transcript for ENSMUSG00000036810: 1843
Updated longest transcript for ENSMUSG00000100254: 145
Updated longest transcript for ENSMUSG00000031176: 1720
Updated longest transcript for ENSMUSG00000061462: 3490
Updated longest transcript for ENSMUSG00000030983: 268
Updated longest transcript for ENSMUSG00000021062: 2283
Updated longest transcript for ENSMUSG00000031290: 2582
Updated longest transcript for ENSMUSG00000019214: 2289
Updated longest transcript for ENSMUSG00000015488: 2111
Updated longest transcript for ENSMUSG00000072780: 9369
Updated longest transcript for ENSMUSG00000024579: 475
Updated longest transcript for ENSMUSG00000052160: 368
Updated longest transcript for ENSMUSG00000038708: 902
Updated longest transcript for ENSMUSG00000051258: 63
Updated longest transcript for ENSMUSG00000020952: 1571
Updated longest transcript for ENSMUSG00000020177: 654
Updated longest transcript for ENSMUSG00000025324: 683
Updated longest transcript for ENSMUSG00000040724: 9401
Updated longest transcript for ENSMUSG00000041117: 871
Updated longest transcript for ENSMUSG00000078324: 151
Updated longest transcript for ENSMUSG00000024780: 4133
Updated longest transcript for ENSMUSG00000023289: 146
Updated longest transcript for ENSMUSG00000026554: 1717
Updated longest transcript for ENSMUSG00000116486: 3492
Updated longest transcript for ENSMUSG00000030703: 18
Updated longest transcript for ENSMUSG00000028799: 1433
Updated longest transcript for ENSMUSG00000035781: 987
Updated longest transcript for ENSMUSG00000068600: 235
Updated longest transcript for ENSMUSG00000006920: 1824
Updated longest transcript for ENSMUSG00000073062: 2891
Updated longest transcript for ENSMUSG00000014846: 343
Updated longest transcript for ENSMUSG00000018589: 1267
Updated longest transcript for ENSMUSG00000032782: 2504
Updated longest transcript for ENSMUSG00000025403: 666
Updated longest transcript for ENSMUSG00000048540: 2431
Updated longest transcript for ENSMUSG00000020312: 1453
Updated longest transcript for ENSMUSG00000008976: 3176
Completed parsing ARE_score_data/3primeUTR_AREscore_GRC38p6_full.txt. Total genes: 21458
Parsing ARE score data from ARE_score_data/3primeUTR_AREscore_GRC39_full.txt...
Warning: No sequence found for ENSMUST00000249497
Warning: No sequence found for ENSMUST00000249279
Updated longest transcript for ENSMUSG00000086587: 386
Warning: No sequence found for ENSMUST00000249374
Updated longest transcript for ENSMUSG00000096574: 1313
Warning: No sequence found for ENSMUST00000249865
Warning: No sequence found for ENSMUST00000249689
Warning: No sequence found for ENSMUST00000247756
Updated longest transcript for ENSMUSG00000079593: 260
Warning: No sequence found for ENSMUST00000249593
Warning: No sequence found for ENSMUST00000249921
Updated longest transcript for ENSMUSG00000078898: 5033
Warning: No sequence found for ENSMUST00000249392
Updated longest transcript for ENSMUSG00000090015: 547
Updated longest transcript for ENSMUSG00000109156: 101
Warning: No sequence found for ENSMUST00000246835
Updated longest transcript for ENSMUSG00000086587: 391
Warning: No sequence found for ENSMUST00000177994
Updated longest transcript for ENSMUSG00000121587: 120
Updated longest transcript for ENSMUSG00000078901: 3470
Warning: No sequence found for ENSMUST00000239509
Updated longest transcript for ENSMUSG00000118664: 1546
Updated longest transcript for ENSMUSG00000078487: 20
Warning: No sequence found for ENSMUST00000239544
Updated longest transcript for ENSMUSG00000079418: 1198
Warning: No sequence found for ENSMUST00000242469
Updated longest transcript for ENSMUSG00000120166: 1356
Warning: No sequence found for ENSMUST00000231152
Updated longest transcript for ENSMUSG00000121584: 247
Warning: No sequence found for ENSMUST00000249298
Updated longest transcript for ENSMUSG00000074529: 532
Warning: No sequence found for ENSMUST00000249872
Warning: No sequence found for ENSMUST00000249889
Updated longest transcript for ENSMUSG00000034354: 1901
Warning: No sequence found for ENSMUST00000249902
Updated longest transcript for ENSMUSG00000020439: 327
Warning: No sequence found for ENSMUST00000249597
Warning: No sequence found for ENSMUST00000249851
Updated longest transcript for ENSMUSG00000121908: 303
Updated longest transcript for ENSMUSG00000079593: 273
Warning: No sequence found for ENSMUST00000249493
Updated longest transcript for ENSMUSG00000091477: 804
Warning: No sequence found for ENSMUST00000249586
Updated longest transcript for ENSMUSG00000050844: 652
Warning: No sequence found for ENSMUST00000105569
Updated longest transcript for ENSMUSG00002076083: 699
Warning: No sequence found for ENSMUST00000249775
Updated longest transcript for ENSMUSG00000050824: 1292
Warning: No sequence found for ENSMUST00000249329
Updated longest transcript for ENSMUSG00000030284: 967
Warning: No sequence found for ENSMUST00000240553
Warning: No sequence found for ENSMUST00000249899
Updated longest transcript for ENSMUSG00000029409: 135
Warning: No sequence found for ENSMUST00000239563
Warning: No sequence found for ENSMUST00000249364
Updated longest transcript for ENSMUSG00000007682: 4872
Warning: No sequence found for ENSMUST00000249742
Warning: No sequence found for ENSMUST00000239547
Updated longest transcript for ENSMUSG00000118669: 1477
Updated longest transcript for ENSMUSG00000118430: 655
Warning: No sequence found for ENSMUST00000249743
Warning: No sequence found for ENSMUST00000249682
Warning: No sequence found for ENSMUST00000249546
Updated longest transcript for ENSMUSG00000095318: 773
Warning: No sequence found for ENSMUST00000249544
Updated longest transcript for ENSMUSG00000095318: 780
Warning: No sequence found for ENSMUST00000249708
Warning: No sequence found for ENSMUST00000249418
Updated longest transcript for ENSMUSG00000072598: 162
Warning: No sequence found for ENSMUST00000240759
Updated longest transcript for ENSMUSG00000094475: 807
Warning: No sequence found for ENSMUST00000249600
Warning: No sequence found for ENSMUST00000249283
Warning: No sequence found for ENSMUST00000249932
Updated longest transcript for ENSMUSG00000095648: 362
Updated longest transcript for ENSMUSG00000036322: 355
Warning: No sequence found for ENSMUST00000249694
Updated longest transcript for ENSMUSG00000015120: 1918
Warning: No sequence found for ENSMUST00000249316
Warning: No sequence found for ENSMUST00000241714
Warning: No sequence found for ENSMUST00000249668
Updated longest transcript for ENSMUSG00000121607: 1326
Updated longest transcript for ENSMUSG00000063171: 435
Warning: No sequence found for ENSMUST00000249501
Warning: No sequence found for ENSMUST00000249924
Updated longest transcript for ENSMUSG00000036779: 2450
Warning: No sequence found for ENSMUST00000240708
Updated longest transcript for ENSMUSG00000114075: 1073
Warning: No sequence found for ENSMUST00000249914
Updated longest transcript for ENSMUSG00000011350: 1365
Warning: No sequence found for ENSMUST00000249333
Warning: No sequence found for ENSMUST00000249701
Updated longest transcript for ENSMUSG00000055114: 403
Warning: No sequence found for ENSMUST00020183810
Warning: No sequence found for ENSMUST00000249337
Updated longest transcript for ENSMUSG00000020633: 983
Warning: No sequence found for ENSMUST00000249702
Warning: No sequence found for ENSMUST00000249378
Updated longest transcript for ENSMUSG00000096793: 1603
Updated longest transcript for ENSMUSG00000079941: 79
Warning: No sequence found for ENSMUST00000249370
Warning: No sequence found for ENSMUST00000249291
Warning: No sequence found for ENSMUST00000249842
Updated longest transcript for ENSMUSG00000015126: 879
Warning: No sequence found for ENSMUST00000240540
Updated longest transcript for ENSMUSG00000113805: 932
Warning: No sequence found for ENSMUST00000249863
Warning: No sequence found for ENSMUST00000249730
Warning: No sequence found for ENSMUST00000249732
Warning: No sequence found for ENSMUST00000249321
Updated longest transcript for ENSMUSG00000074529: 600
Warning: No sequence found for ENSMUST00000249663
Updated longest transcript for ENSMUSG00000121605: 119
Warning: No sequence found for ENSMUST00000249068
Warning: No sequence found for ENSMUST00000249287
Updated longest transcript for ENSMUSG00000022253: 2534
Updated longest transcript for ENSMUSG00000024176: 1673
Warning: No sequence found for ENSMUST00000249257
Updated longest transcript for ENSMUSG00000075023: 483
Warning: No sequence found for ENSMUST00000249923
Updated longest transcript for ENSMUSG00000073001: 107
Warning: No sequence found for ENSMUST00000249750
Warning: No sequence found for ENSMUST00000249695
Updated longest transcript for ENSMUSG00000063277: 1139
Warning: No sequence found for ENSMUST00000249885
Updated longest transcript for ENSMUSG00000062691: 1072
Warning: No sequence found for ENSMUST00000240548
Warning: No sequence found for ENSMUST00000249876
Warning: No sequence found for ENSMUST00000240547
Updated longest transcript for ENSMUSG00000000386: 1647
Warning: No sequence found for ENSMUST00000249221
Updated longest transcript for ENSMUSG00000094091: 344
Warning: No sequence found for ENSMUST00000249773
Warning: No sequence found for ENSMUST00000247757
Warning: No sequence found for ENSMUST00000240713
Updated longest transcript for ENSMUSG00000092335: 2252
Warning: No sequence found for ENSMUST00000249607
Updated longest transcript for ENSMUSG00000094132: 1057
Updated longest transcript for ENSMUSG00000116376: 123
Warning: No sequence found for ENSMUST00000240719
Warning: No sequence found for ENSMUST00000247219
Warning: No sequence found for ENSMUST00000249267
Updated longest transcript for ENSMUSG00000073448: 444
Warning: No sequence found for ENSMUST00010181660
Updated longest transcript for ENSMUSG00001074846: 53
Warning: No sequence found for ENSMUST00000249683
Warning: No sequence found for ENSMUST00000249361
Updated longest transcript for ENSMUSG00000095648: 462
Warning: No sequence found for ENSMUST00020183806
Updated longest transcript for ENSMUSG00000062691: 1074
Warning: No sequence found for ENSMUST00010126032
Updated longest transcript for ENSMUSG00001074846: 460
Warning: No sequence found for ENSMUST00000240709
Warning: No sequence found for ENSMUST00000249302
Updated longest transcript for ENSMUSG00000015120: 1919
Warning: No sequence found for ENSMUST00000249933
Updated longest transcript for ENSMUSG00000118415: 2396
Warning: No sequence found for ENSMUST00000249918
Warning: No sequence found for ENSMUST00000240546
Warning: No sequence found for ENSMUST00000249434
Warning: No sequence found for ENSMUST00000239500
Updated longest transcript for ENSMUSG00000118661: 589
Warning: No sequence found for ENSMUST00000249858
Updated longest transcript for ENSMUSG00000093806: 342
Warning: No sequence found for ENSMUST00000249891
Warning: No sequence found for ENSMUST00000249388
Updated longest transcript for ENSMUSG00000121596: 1420
Warning: No sequence found for ENSMUST00000239562
Warning: No sequence found for ENSMUST00000248980
Warning: No sequence found for ENSMUST00000248072
Warning: No sequence found for ENSMUST00000240716
Warning: No sequence found for ENSMUST00000239529
Updated longest transcript for ENSMUSG00000118668: 190
Warning: No sequence found for ENSMUST00000249864
Warning: No sequence found for ENSMUST00000249379
Warning: No sequence found for ENSMUST00000249747
Warning: No sequence found for ENSMUST00000240758
Warning: No sequence found for ENSMUST00000247844
Warning: No sequence found for ENSMUST00000249247
Warning: No sequence found for ENSMUST00000249919
Warning: No sequence found for ENSMUST00000239519
Updated longest transcript for ENSMUSG00000073791: 1655
Warning: No sequence found for ENSMUST00000249613
Warning: No sequence found for ENSMUST00000249705
Warning: No sequence found for ENSMUST00000249277
Updated longest transcript for ENSMUSG00000094932: 869
Warning: No sequence found for ENSMUST00000239551
Warning: No sequence found for ENSMUST00020183325
Updated longest transcript for ENSMUSG00002076083: 2117
Warning: No sequence found for ENSMUST00000249274
Updated longest transcript for ENSMUSG00000028883: 3217
Warning: No sequence found for ENSMUST00000240706
Warning: No sequence found for ENSMUST00000249584
Updated longest transcript for ENSMUSG00000096619: 567
Warning: No sequence found for ENSMUST00000249866
Updated longest transcript for ENSMUSG00000095799: 928
Updated longest transcript for ENSMUSG00000024171: 206
Warning: No sequence found for ENSMUST00000249241
Updated longest transcript for ENSMUSG00000121570: 8
Warning: No sequence found for ENSMUST00000239536
Updated longest transcript for ENSMUSG00000118669: 1482
Warning: No sequence found for ENSMUST00000249913
Updated longest transcript for ENSMUSG00000029447: 551
Warning: No sequence found for ENSMUST00000249862
Updated longest transcript for ENSMUSG00000008658: 1836
Updated longest transcript for ENSMUSG00000118516: 2774
Warning: No sequence found for ENSMUST00000249860
Updated longest transcript for ENSMUSG00000079164: 691
Updated longest transcript for ENSMUSG00000114591: 729
Warning: No sequence found for ENSMUST00000249368
Warning: No sequence found for ENSMUST00000189062
Updated longest transcript for ENSMUSG00000121918: 1338
Warning: No sequence found for ENSMUST00000249868
Updated longest transcript for ENSMUSG00000058183: 1686
Updated longest transcript for ENSMUSG00000015451: 105
Warning: No sequence found for ENSMUST00000249699
Warning: No sequence found for ENSMUST00000239534
Warning: No sequence found for ENSMUST00000249567
Updated longest transcript for ENSMUSG00000091568: 1136
Updated longest transcript for ENSMUSG00000074521: 3609
Warning: No sequence found for ENSMUST00000249348
Warning: No sequence found for ENSMUST00000249925
Warning: No sequence found for ENSMUST00000249713
Warning: No sequence found for ENSMUST00000249620
Warning: No sequence found for ENSMUST00000249908
Updated longest transcript for ENSMUSG00000053253: 1768
Warning: No sequence found for ENSMUST00020183803
Warning: No sequence found for ENSMUST00000249608
Warning: No sequence found for ENSMUST00000249313
Warning: No sequence found for ENSMUST00000249494
Updated longest transcript for ENSMUSG00000091477: 811
Warning: No sequence found for ENSMUST00000249935
Updated longest transcript for ENSMUSG00000121920: 264
Warning: No sequence found for ENSMUST00000249827
Warning: No sequence found for ENSMUST00000249455
Updated longest transcript for ENSMUSG00000017688: 2624
Updated longest transcript for ENSMUSG00000049133: 598
Warning: No sequence found for ENSMUST00000185223
Updated longest transcript for ENSMUSG00000023259: 1694
Warning: No sequence found for ENSMUST00000240551
Updated longest transcript for ENSMUSG00000027157: 1215
Warning: No sequence found for ENSMUST00000239516
Warning: No sequence found for ENSMUST00000249300
Warning: No sequence found for ENSMUST00000249293
Updated longest transcript for ENSMUSG00000095474: 2391
Warning: No sequence found for ENSMUST00000249372
Updated longest transcript for ENSMUSG00000091110: 1688
Warning: No sequence found for ENSMUST00000240589
Warning: No sequence found for ENSMUST00000249502
Updated longest transcript for ENSMUSG00000004677: 1255
Warning: No sequence found for ENSMUST00000249320
Warning: No sequence found for ENSMUST00000249395
Warning: No sequence found for ENSMUST00000249363
Warning: No sequence found for ENSMUST00000249366
Warning: No sequence found for ENSMUST00000249308
Warning: No sequence found for ENSMUST00000249898
Warning: No sequence found for ENSMUST00000249290
Updated longest transcript for ENSMUSG00000024942: 817
Warning: No sequence found for ENSMUST00000249765
Warning: No sequence found for ENSMUST00000249458
Warning: No sequence found for ENSMUST00000249304
Updated longest transcript for ENSMUSG00000095180: 117
Warning: No sequence found for ENSMUST00000249389
Warning: No sequence found for ENSMUST00000249621
Updated longest transcript for ENSMUSG00000032846: 1818
Warning: No sequence found for ENSMUST00000249761
Updated longest transcript for ENSMUSG00000121905: 115
Warning: No sequence found for ENSMUST00000240714
Warning: No sequence found for ENSMUST00000247843
Warning: No sequence found for ENSMUST00000249431
Warning: No sequence found for ENSMUST00000249833
Warning: No sequence found for ENSMUST00000249697
Warning: No sequence found for ENSMUST00000249612
Warning: No sequence found for ENSMUST00000247868
Updated longest transcript for ENSMUSG00000026321: 3117
Updated longest transcript for ENSMUSG00000036073: 429
Warning: No sequence found for ENSMUST00000239521
Warning: No sequence found for ENSMUST00000249631
Updated longest transcript for ENSMUSG00000110218: 1691
Warning: No sequence found for ENSMUST00000249401
Updated longest transcript for ENSMUSG00000114582: 729
Warning: No sequence found for ENSMUST00000249849
Updated longest transcript for ENSMUSG00000082226: 368
Warning: No sequence found for ENSMUST00000153071
Updated longest transcript for ENSMUSG00000121906: 337
Warning: No sequence found for ENSMUST00000240549
Warning: No sequence found for ENSMUST00000249382
Updated longest transcript for ENSMUSG00000095195: 1134
Warning: No sequence found for ENSMUST00000249299
Warning: No sequence found for ENSMUST00000239541
Updated longest transcript for ENSMUSG00000073991: 35
Warning: No sequence found for ENSMUST00000249276
Warning: No sequence found for ENSMUST00000249305
Warning: No sequence found for ENSMUST00000249762
Updated longest transcript for ENSMUSG00000024175: 859
Warning: No sequence found for ENSMUST00000249585
Warning: No sequence found for ENSMUST00000075040
Updated longest transcript for ENSMUSG00000121906: 1150
Warning: No sequence found for ENSMUST00000239561
Warning: No sequence found for ENSMUST00000249110
Warning: No sequence found for ENSMUST00000249691
Warning: No sequence found for ENSMUST00000249376
Warning: No sequence found for ENSMUST00000249763
Warning: No sequence found for ENSMUST00000249602
Warning: No sequence found for ENSMUST00000249895
Warning: No sequence found for ENSMUST00000249436
Warning: No sequence found for ENSMUST00000249617
Warning: No sequence found for ENSMUST00000240715
Warning: No sequence found for ENSMUST00000249703
Updated longest transcript for ENSMUSG00000026285: 1003
Warning: No sequence found for ENSMUST00000249551
Warning: No sequence found for ENSMUST00000249355
Warning: No sequence found for ENSMUST00000249594
Warning: No sequence found for ENSMUST00000249384
Updated longest transcript for ENSMUSG00000031838: 201
Warning: No sequence found for ENSMUST00000249475
Updated longest transcript for ENSMUSG00000069355: 551
Warning: No sequence found for ENSMUST00000249879
Updated longest transcript for ENSMUSG00000031714: 1895
Warning: No sequence found for ENSMUST00000249867
Warning: No sequence found for ENSMUST00000239525
Updated longest transcript for ENSMUSG00000118667: 712
Warning: No sequence found for ENSMUST00000249256
Warning: No sequence found for ENSMUST00000249285
Updated longest transcript for ENSMUSG00000092572: 2199
Warning: No sequence found for ENSMUST00000240702
Updated longest transcript for ENSMUSG00000013083: 156
Warning: No sequence found for ENSMUST00000249433
Updated longest transcript for ENSMUSG00000096803: 366
Warning: No sequence found for ENSMUST00000249598
Warning: No sequence found for ENSMUST00000249258
Updated longest transcript for ENSMUSG00000121585: 123
Warning: No sequence found for ENSMUST00000249319
Updated longest transcript for ENSMUSG00000095545: 7392
Warning: No sequence found for ENSMUST00000249604
Updated longest transcript for ENSMUSG00000094132: 1059
Updated longest transcript for ENSMUSG00000085549: 222
Warning: No sequence found for ENSMUST00000249733
Warning: No sequence found for ENSMUST00000239545
Updated longest transcript for ENSMUSG00000118670: 184
Warning: No sequence found for ENSMUST00000249934
Updated longest transcript for ENSMUSG00000113971: 929
Updated longest transcript for ENSMUSG00000034039: 178
Warning: No sequence found for ENSMUST00000249595
Warning: No sequence found for ENSMUST00000247846
Warning: No sequence found for ENSMUST00000249432
Warning: No sequence found for ENSMUST00000249120
Updated longest transcript for ENSMUSG00000086158: 173
Warning: No sequence found for ENSMUST00000249296
Warning: No sequence found for ENSMUST00000249312
Warning: No sequence found for ENSMUST00000249097
Updated longest transcript for ENSMUSG00000073991: 399
Warning: No sequence found for ENSMUST00000249656
Updated longest transcript for ENSMUSG00000020894: 2092
Warning: No sequence found for ENSMUST00000239552
Updated longest transcript for ENSMUSG00000118671: 822
Updated longest transcript for ENSMUSG00000024942: 818
Updated longest transcript for ENSMUSG00000053961: 162
Warning: No sequence found for ENSMUST00000249263
Warning: No sequence found for ENSMUST00000249073
Warning: No sequence found for ENSMUST00000249615
Warning: No sequence found for ENSMUST00000249233
Updated longest transcript for ENSMUSG00000024729: 123
Warning: No sequence found for ENSMUST00000249622
Warning: No sequence found for ENSMUST00000249237
Updated longest transcript for ENSMUSG00000024729: 688
Warning: No sequence found for ENSMUST00000249828
Warning: No sequence found for ENSMUST00000249774
Updated longest transcript for ENSMUSG00000078867: 2802
Warning: No sequence found for ENSMUST00000249476
Updated longest transcript for ENSMUSG00000056938: 3466
Warning: No sequence found for ENSMUST00000249861
Updated longest transcript for ENSMUSG00000072762: 1238
Warning: No sequence found for ENSMUST00000249870
Updated longest transcript for ENSMUSG00000090093: 1072
Warning: No sequence found for ENSMUST00000249706
Warning: No sequence found for ENSMUST00000249840
Warning: No sequence found for ENSMUST00000249338
Updated longest transcript for ENSMUSG00000020633: 990
Updated longest transcript for ENSMUSG00000070686: 1493
Warning: No sequence found for ENSMUST00000249231
Warning: No sequence found for ENSMUST00000249552
Warning: No sequence found for ENSMUST00000249457
Updated longest transcript for ENSMUSG00000094021: 647
Warning: No sequence found for ENSMUST00000239523
Updated longest transcript for ENSMUSG00000095180: 120
Warning: No sequence found for ENSMUST00000249909
Updated longest transcript for ENSMUSG00000096049: 922
Warning: No sequence found for ENSMUST00000249232
Warning: No sequence found for ENSMUST00000249242
Updated longest transcript for ENSMUSG00000121571: 662
Warning: No sequence found for ENSMUST00000249230
Warning: No sequence found for ENSMUST00000249369
Warning: No sequence found for ENSMUST00000249739
Warning: No sequence found for ENSMUST00000249538
Updated longest transcript for ENSMUSG00000094157: 759
Warning: No sequence found for ENSMUST00000249286
Warning: No sequence found for ENSMUST00000249549
Warning: No sequence found for ENSMUST00000240707
Warning: No sequence found for ENSMUST00000249259
Updated longest transcript for ENSMUSG00000100679: 1651
Warning: No sequence found for ENSMUST00000249720
Warning: No sequence found for ENSMUST00000249474
Updated longest transcript for ENSMUSG00000091617: 1318
Warning: No sequence found for ENSMUST00000249704
Warning: No sequence found for ENSMUST00000249331
Warning: No sequence found for ENSMUST00000247847
Updated longest transcript for ENSMUSG00000107539: 492
Warning: No sequence found for ENSMUST00010126065
Warning: No sequence found for ENSMUST00000249547
Warning: No sequence found for ENSMUST00000249603
Warning: No sequence found for ENSMUST00000249235
Warning: No sequence found for ENSMUST00000249317
Updated longest transcript for ENSMUSG00000095936: 1112
Updated longest transcript for ENSMUSG00000017466: 2638
Warning: No sequence found for ENSMUST00000249871
Warning: No sequence found for ENSMUST00000249377
Warning: No sequence found for ENSMUST00000249661
Warning: No sequence found for ENSMUST00000249303
Warning: No sequence found for ENSMUST00000239527
Updated longest transcript for ENSMUSG00000118668: 729
Warning: No sequence found for ENSMUST00000249495
Warning: No sequence found for ENSMUST00000247845
Warning: No sequence found for ENSMUST00000240711
Warning: No sequence found for ENSMUST00020183804
Warning: No sequence found for ENSMUST00000240710
Warning: No sequence found for ENSMUST00000240543
Warning: No sequence found for ENSMUST00000249043
Updated longest transcript for ENSMUSG00000094443: 115
Warning: No sequence found for ENSMUST00000212885
Updated longest transcript for ENSMUSG00000121369: 183
Updated longest transcript for ENSMUSG00000078510: 577
Warning: No sequence found for ENSMUST00000249244
Updated longest transcript for ENSMUSG00000056399: 166
Warning: No sequence found for ENSMUST00000249109
Warning: No sequence found for ENSMUST00000249915
Updated longest transcript for ENSMUSG00000060913: 1105
Warning: No sequence found for ENSMUST00000249721
Warning: No sequence found for ENSMUST00000249881
Warning: No sequence found for ENSMUST00000240539
Warning: No sequence found for ENSMUST00000247866
Warning: No sequence found for ENSMUST00000249074
Updated longest transcript for ENSMUSG00000073085: 206
Warning: No sequence found for ENSMUST00000249498
Warning: No sequence found for ENSMUST00000249890
Warning: No sequence found for ENSMUST00000249901
Warning: No sequence found for ENSMUST00000249671
Warning: No sequence found for ENSMUST00000249907
Warning: No sequence found for ENSMUST00000249681
Warning: No sequence found for ENSMUST00000249469
Warning: No sequence found for ENSMUST00000249236
Warning: No sequence found for ENSMUST00000249289
Warning: No sequence found for ENSMUST00000249133
Warning: No sequence found for ENSMUST00000249712
Warning: No sequence found for ENSMUST00000249666
Updated longest transcript for ENSMUSG00000121606: 216
Warning: No sequence found for ENSMUST00000249568
Warning: No sequence found for ENSMUST00000249606
Warning: No sequence found for ENSMUST00000249096
Warning: No sequence found for ENSMUST00000249278
Warning: No sequence found for ENSMUST00000249234
Updated longest transcript for ENSMUSG00000114694: 729
Warning: No sequence found for ENSMUST00000249288
Warning: No sequence found for ENSMUST00000239538
Warning: No sequence found for ENSMUST00000239514
Updated longest transcript for ENSMUSG00000118665: 3265
Warning: No sequence found for ENSMUST00000249931
Warning: No sequence found for ENSMUST00000249619
Warning: No sequence found for ENSMUST00010181659
Warning: No sequence found for ENSMUST00000249078
Updated longest transcript for ENSMUSG00000090379: 768
Updated longest transcript for ENSMUSG00000048424: 2330
Warning: No sequence found for ENSMUST00000249387
Warning: No sequence found for ENSMUST00000249900
Updated longest transcript for ENSMUSG00000041361: 2652
Warning: No sequence found for ENSMUST00000248712
Updated longest transcript for ENSMUSG00000121513: 4113
Warning: No sequence found for ENSMUST00000240542
Warning: No sequence found for ENSMUST00000249897
Warning: No sequence found for ENSMUST00000249496
Warning: No sequence found for ENSMUST00000249926
Warning: No sequence found for ENSMUST00000249292
Updated longest transcript for ENSMUSG00000118483: 2356
Warning: No sequence found for ENSMUST00000249835
Warning: No sequence found for ENSMUST00000249917
Warning: No sequence found for ENSMUST00000239513
Updated longest transcript for ENSMUSG00000078898: 7392
Warning: No sequence found for ENSMUST00000249269
Updated longest transcript for ENSMUSG00000073448: 683
Updated longest transcript for ENSMUSG00000060244: 443
Warning: No sequence found for ENSMUST00000249687
Updated longest transcript for ENSMUSG00000015120: 2030
Warning: No sequence found for ENSMUST00000249837
Warning: No sequence found for ENSMUST00000177359
Warning: No sequence found for ENSMUST00000249859
Updated longest transcript for ENSMUSG00000036879: 2105
Warning: No sequence found for ENSMUST00000249875
Warning: No sequence found for ENSMUST00000249735
Updated longest transcript for ENSMUSG00000033825: 801
Updated longest transcript for ENSMUSG00000023572: 512
Warning: No sequence found for ENSMUST00000249692
Warning: No sequence found for ENSMUST00000249610
Warning: No sequence found for ENSMUST00000249834
Warning: No sequence found for ENSMUST00000249281
Warning: No sequence found for ENSMUST00000239554
Updated longest transcript for ENSMUSG00000118672: 205
Warning: No sequence found for ENSMUST00000249737
Warning: No sequence found for ENSMUST00000249381
Warning: No sequence found for ENSMUST00000249250
Updated longest transcript for ENSMUSG00000121577: 666
Warning: No sequence found for ENSMUST00000249831
Warning: No sequence found for ENSMUST00000239506
Updated longest transcript for ENSMUSG00000118663: 56
Warning: No sequence found for ENSMUST00000249310
Warning: No sequence found for ENSMUST00000249350
Warning: No sequence found for ENSMUST00000248112
Updated longest transcript for ENSMUSG00000091122: 770
Warning: No sequence found for ENSMUST00020183129
Warning: No sequence found for ENSMUST00000249280
Warning: No sequence found for ENSMUST00000240721
Warning: No sequence found for ENSMUST00000249611
Updated longest transcript for ENSMUSG00000094132: 1308
Warning: No sequence found for ENSMUST00000249740
Updated longest transcript for ENSMUSG00000028108: 94
Updated longest transcript for ENSMUSG00000049608: 2557
Warning: No sequence found for ENSMUST00000249841
Warning: No sequence found for ENSMUST00000249772
Updated longest transcript for ENSMUSG00000064043: 2727
Warning: No sequence found for ENSMUST00000249839
Warning: No sequence found for ENSMUST00000249892
Warning: No sequence found for ENSMUST00000249327
Warning: No sequence found for ENSMUST00000239508
Warning: No sequence found for ENSMUST00000247875
Warning: No sequence found for ENSMUST00000249371
Updated longest transcript for ENSMUSG00000025716: 309
Warning: No sequence found for ENSMUST00000249284
Warning: No sequence found for ENSMUST00000249295
Warning: No sequence found for ENSMUST00000249545
Updated longest transcript for ENSMUSG00000095318: 1159
Warning: No sequence found for ENSMUST00000249471
Warning: No sequence found for ENSMUST00000249887
Warning: No sequence found for ENSMUST00000249764
Warning: No sequence found for ENSMUST00000248537
Updated longest transcript for ENSMUSG00000121513: 4168
Warning: No sequence found for ENSMUST00000249380
Warning: No sequence found for ENSMUST00000249477
Warning: No sequence found for ENSMUST00000249282
Warning: No sequence found for ENSMUST00000249403
Warning: No sequence found for ENSMUST00000249470
Warning: No sequence found for ENSMUST00000249880
Warning: No sequence found for ENSMUST00000249711
Warning: No sequence found for ENSMUST00000249228
Updated longest transcript for ENSMUSG00000055555: 586
Warning: No sequence found for ENSMUST00020183805
Warning: No sequence found for ENSMUST00000249684
Warning: No sequence found for ENSMUST00000239510
Warning: No sequence found for ENSMUST00000249709
Warning: No sequence found for ENSMUST00000249266
Warning: No sequence found for ENSMUST00000249349
Updated longest transcript for ENSMUSG00000029673: 2103
Warning: No sequence found for ENSMUST00000247867
Warning: No sequence found for ENSMUST00000249294
Updated longest transcript for ENSMUSG00000093701: 711
Warning: No sequence found for ENSMUST00000249306
Updated longest transcript for ENSMUSG00000105888: 126
Warning: No sequence found for ENSMUST00000240717
Updated longest transcript for ENSMUSG00000080152: 316
Warning: No sequence found for ENSMUST00000066677
Warning: No sequence found for ENSMUST00000249710
Warning: No sequence found for ENSMUST00000249857
Warning: No sequence found for ENSMUST00000249698
Warning: No sequence found for ENSMUST00000240722
Warning: No sequence found for ENSMUST00000249894
Warning: No sequence found for ENSMUST00000240671
Warning: No sequence found for ENSMUST00000249693
Warning: No sequence found for ENSMUST00000249669
Updated longest transcript for ENSMUSG00000121607: 2395
Warning: No sequence found for ENSMUST00000239535
Warning: No sequence found for ENSMUST00000239533
Warning: No sequence found for ENSMUST00000249850
Updated longest transcript for ENSMUSG00000068686: 986
Warning: No sequence found for ENSMUST00000249273
Warning: No sequence found for ENSMUST00000249307
Warning: No sequence found for ENSMUST00000249719
Warning: No sequence found for ENSMUST00000239501
Updated longest transcript for ENSMUSG00000118662: 506
Warning: No sequence found for ENSMUST00000249751
Warning: No sequence found for ENSMUST00000249700
Warning: No sequence found for ENSMUST00000249728
Warning: No sequence found for ENSMUST00000239548
Warning: No sequence found for ENSMUST00000240720
Warning: No sequence found for ENSMUST00000249729
Updated longest transcript for ENSMUSG00000002111: 619
Warning: No sequence found for ENSMUST00000249565
Warning: No sequence found for ENSMUST00000249375
Warning: No sequence found for ENSMUST00000249330
Updated longest transcript for ENSMUSG00000095909: 1303
Updated longest transcript for ENSMUSG00000073650: 1753
Warning: No sequence found for ENSMUST00000249328
Updated longest transcript for ENSMUSG00000099216: 646
Warning: No sequence found for ENSMUST00000249248
Updated longest transcript for ENSMUSG00000121575: 107
Warning: No sequence found for ENSMUST00000249601
Warning: No sequence found for ENSMUST00000249678
Warning: No sequence found for ENSMUST00000249566
Updated longest transcript for ENSMUSG00000063235: 2845
Warning: No sequence found for ENSMUST00000249357
Warning: No sequence found for ENSMUST00000249605
Warning: No sequence found for ENSMUST00000249878
Updated longest transcript for ENSMUSG00000091601: 1341
Updated longest transcript for ENSMUSG00000090093: 1792
Warning: No sequence found for ENSMUST00000249591
Warning: No sequence found for ENSMUST00000249830
Warning: No sequence found for ENSMUST00000249609
Updated longest transcript for ENSMUSG00000015869: 1096
Warning: No sequence found for ENSMUST00000249540
Updated longest transcript for ENSMUSG00000094157: 1820
Warning: No sequence found for ENSMUST00000249367
Warning: No sequence found for ENSMUST00000249301
Warning: No sequence found for ENSMUST00000249718
Updated longest transcript for ENSMUSG00000079697: 178
Updated longest transcript for ENSMUSG00000051146: 958
Warning: No sequence found for ENSMUST00000249896
Warning: No sequence found for ENSMUST00000249456
Warning: No sequence found for ENSMUST00000240712
Warning: No sequence found for ENSMUST00000249275
Warning: No sequence found for ENSMUST00000249667
Warning: No sequence found for ENSMUST00000249326
Updated longest transcript for ENSMUSG00000092335: 3208
Warning: No sequence found for ENSMUST00000249760
Updated longest transcript for ENSMUSG00000121904: 145
Warning: No sequence found for ENSMUST00000249268
Updated longest transcript for ENSMUSG00000036073: 645
Warning: No sequence found for ENSMUST00000249383
Warning: No sequence found for ENSMUST00000249888
Warning: No sequence found for ENSMUST00000249884
Updated longest transcript for ENSMUSG00000034354: 2166
Updated longest transcript for ENSMUSG00000095029: 154
Warning: No sequence found for ENSMUST00000249314
Warning: No sequence found for ENSMUST00000249664
Updated longest transcript for ENSMUSG00000121605: 121
Updated longest transcript for ENSMUSG00000113201: 931
Warning: No sequence found for ENSMUST00000249744
Updated longest transcript for ENSMUSG00000072258: 848
Warning: No sequence found for ENSMUST00000249893
Updated longest transcript for ENSMUSG00000095648: 1040
Warning: No sequence found for ENSMUST00000249309
Warning: No sequence found for ENSMUST00000249829
Warning: No sequence found for ENSMUST00000249745
Warning: No sequence found for ENSMUST00000249500
Warning: No sequence found for ENSMUST00000249920
Warning: No sequence found for ENSMUST00000249359
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Completed parsing ARE_score_data/3primeUTR_AREscore_GRC39_full.txt. Total genes: 21540
Completed parsing ARE score data and calculating motif counts.
Updating 6 `upregulated` files in upregulated_genes...
Processed 1/6 files. Current file: BM_upregulated_annotated.csv
Processed 2/6 files. Current file: Spleen_upregulated_annotated.csv
Processed 3/6 files. Current file: Ovary_upregulated_annotated.csv
Processed 4/6 files. Current file: Lung_upregulated_annotated.csv
Processed 5/6 files. Current file: Liver_upregulated_annotated.csv
Processed 6/6 files. Current file: Kidney_upregulated_annotated.csv
Completed updating files in upregulated_genes.
Updating 6 `unregulated` files in unregulated_genes...
Processed 1/6 files. Current file: Kidney_unregulated_annotated.csv
Processed 2/6 files. Current file: Spleen_unregulated_annotated.csv
Processed 3/6 files. Current file: Lung_unregulated_annotated.csv
Processed 4/6 files. Current file: BM_unregulated_annotated.csv
Processed 5/6 files. Current file: Ovary_unregulated_annotated.csv
Processed 6/6 files. Current file: Liver_unregulated_annotated.csv
Completed updating files in unregulated_genes.
Updating 6 `downregulated` files in downregulated_genes...
Processed 1/6 files. Current file: Spleen_downregulated_annotated.csv
Processed 2/6 files. Current file: BM_downregulated_annotated.csv
Processed 3/6 files. Current file: Ovary_downregulated_annotated.csv
Processed 4/6 files. Current file: Lung_downregulated_annotated.csv
Processed 5/6 files. Current file: Kidney_downregulated_annotated.csv
Processed 6/6 files. Current file: Liver_downregulated_annotated.csv
Completed updating files in downregulated_genes.
All files have been updated successfully.

Below will be my first iteration of a upset plot wiht two panes includng ARE score information and overlapping set information

In [4]:
import os
import pandas as pd
import matplotlib.pyplot as plt
import seaborn as sns
from upsetplot import UpSet, from_indicators

# Function to create the combined UpSet and Violin plot
def create_upset_violin(data, title, output_file, color):
    indicator_cols = ['BM', 'Spleen', 'Ovary', 'Liver', 'Lung', 'Kidney']
    
    # Ensure all indicator columns are boolean type
    data[indicator_cols] = data[indicator_cols].astype(bool)

    # Filter out rows where ARE_score is NaN
    data = data.dropna(subset=['ARE_score'])
    data['ARE_score'] = pd.to_numeric(data['ARE_score'], errors='coerce')

    # Create the UpSet plot data
    upset_data = from_indicators(indicator_cols, data)

    # Initialize UpSet plot
    upset = UpSet(upset_data, subset_size='count', sort_by='degree', facecolor=color)

    # Add the violin plots for ARE_score values
    upset.add_catplot(
        value='ARE_score', 
        kind='violin', 
        elements=2, 
        color=color
    )

    # Plot UpSet with catplots
    upset.plot()
    plt.suptitle(f'{title} Genes with ARE Score Distribution')

    # Save the combined plot
    plt.savefig(output_file)
    plt.show()

# Load and concatenate data from all updated files in the specified directory
def load_and_concat_data(directory):
    files = [os.path.join(directory, file) for file in os.listdir(directory) if file.endswith('_annotated_updated.csv')]
    data = pd.concat((pd.read_csv(file) for file in files), ignore_index=True)
    return data

# Load upregulated and downregulated data
up_data = load_and_concat_data("upregulated_genes_updated")
down_data = load_and_concat_data("downregulated_genes_updated")

# Create UpSet plots with violin plots
create_upset_violin(up_data, 'Upregulated', 'upregulated_genes_upset_violin.png', 'red')
create_upset_violin(down_data, 'Downregulated', 'downregulated_genes_upset_violin.png', 'blue')
/tmp/ipykernel_2823251/4268302770.py:16: SettingWithCopyWarning: 
A value is trying to be set on a copy of a slice from a DataFrame.
Try using .loc[row_indexer,col_indexer] = value instead

See the caveats in the documentation: https://pandas.pydata.org/pandas-docs/stable/user_guide/indexing.html#returning-a-view-versus-a-copy
  data['ARE_score'] = pd.to_numeric(data['ARE_score'], errors='coerce')
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:795: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linewidth"].fillna(1, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:796: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["facecolor"].fillna(self._facecolor, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:797: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["edgecolor"].fillna(styles["facecolor"], inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:798: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linestyle"].fillna("solid", inplace=True)
No description has been provided for this image
/tmp/ipykernel_2823251/4268302770.py:16: SettingWithCopyWarning: 
A value is trying to be set on a copy of a slice from a DataFrame.
Try using .loc[row_indexer,col_indexer] = value instead

See the caveats in the documentation: https://pandas.pydata.org/pandas-docs/stable/user_guide/indexing.html#returning-a-view-versus-a-copy
  data['ARE_score'] = pd.to_numeric(data['ARE_score'], errors='coerce')
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:795: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linewidth"].fillna(1, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:796: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["facecolor"].fillna(self._facecolor, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:797: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["edgecolor"].fillna(styles["facecolor"], inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:798: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linestyle"].fillna("solid", inplace=True)
No description has been provided for this image
In [5]:
import os
import pandas as pd

# Helper function to load and concatenate data from all updated files in the specified directory
def load_and_concat_data(directory):
    files = [os.path.join(directory, file) for file in os.listdir(directory) if file.endswith('_annotated_updated.csv')]
    data = pd.concat((pd.read_csv(file) for file in files), ignore_index=True)
    return data

# Load upregulated data
upregulated_path = "upregulated_genes_updated"
up_data = load_and_concat_data(upregulated_path)

# Ensure all indicator columns are boolean type
indicator_cols = ['BM', 'Spleen', 'Ovary', 'Liver', 'Lung', 'Kidney']
up_data[indicator_cols] = up_data[indicator_cols].astype(bool)

# Filter to only include protein_coding genes
up_data = up_data[up_data['gene_biotype'] == 'protein_coding']

# Calculate the sum of binary indicators for each gene
up_data['sum_tissues'] = up_data[indicator_cols].sum(axis=1)

# List genes that are upregulated in 5 or more tissues
upregulated_5_or_more = up_data[up_data['sum_tissues'] >= 5]

# Display the list of gene IDs
upregulated_5_or_more_ids = upregulated_5_or_more['ID']
print(upregulated_5_or_more_ids.tolist())
['ENSMUSG00000035352', 'ENSMUSG00000041481', 'ENSMUSG00000091694', 'ENSMUSG00000035352', 'ENSMUSG00000041481', 'ENSMUSG00000091694', 'ENSMUSG00000091694', 'ENSMUSG00000041481', 'ENSMUSG00000035352', 'ENSMUSG00000091694', 'ENSMUSG00000035352', 'ENSMUSG00000041481', 'ENSMUSG00000091694', 'ENSMUSG00000035352', 'ENSMUSG00000041481', 'ENSMUSG00000091694']

Now I make the same plot again but this time I added strip plots instead of box and whisker plots

In [6]:
import os
import pandas as pd
import matplotlib.pyplot as plt
from upsetplot import UpSet, from_indicators

# Function to create the combined UpSet and Violin plot
def create_upset_violin(data, title, output_file, color):
    indicator_cols = ['BM', 'Spleen', 'Ovary', 'Liver', 'Lung', 'Kidney']
    # Ensure all indicator columns are boolean type
    data[indicator_cols] = data[indicator_cols].astype(bool)
    # Filter out rows where ARE_score is NaN and Gene.biotype is not 'protein_coding'
    data = data[data['gene_biotype'] == 'protein_coding']
    data = data.dropna(subset=['ARE_score'])
    data['ARE_score'] = pd.to_numeric(data['ARE_score'], errors='coerce')
    
    # Group by gene ID and aggregate the Boolean indicators
    data_agg = data.groupby('ID')[indicator_cols + ['ARE_score']].max()
    
    # Ensure every row is treated as unique for UpSet plot
    data_agg = data_agg.reset_index()

    # Create the UpSet plot data
    upset_data = from_indicators(indicator_cols, data_agg)
    
    # Initialize UpSet plot with desired formatting
    upset = UpSet(
        upset_data,
        subset_size='count',
        sort_by='-degree',
        facecolor=color,
        show_counts=True,
        max_subset_size=30 # Limiting the large number of insignificant intersections
    )
    
    # Add the violin plots for ARE_score values
    upset.add_catplot(
        value='ARE_score',
        kind='strip',
        elements=2,
        color=color
    )
    
    # Plot UpSet with catplots
    upset.plot()
    
    # Add title for the combined plot
    plt.suptitle(f'{title} Genes with ARE Score Distribution')
    
    # Save the combined plot
    plt.savefig(output_file)
    plt.show()

# Load and concatenate data from all updated files in the specified directory
def load_and_concat_data(directory):
    files = [os.path.join(directory, file) for file in os.listdir(directory) if file.endswith('_annotated_updated.csv')]
    data = pd.concat((pd.read_csv(file) for file in files), ignore_index=True)
    return data

# Load upregulated and downregulated data
up_data = load_and_concat_data("upregulated_genes_updated")
down_data = load_and_concat_data("downregulated_genes_updated")

# Create UpSet plots with violin plots
create_upset_violin(up_data, 'Upregulated', 'upregulated_genes_upset_violin.png', 'red')
create_upset_violin(down_data, 'Downregulated', 'downregulated_genes_upset_violin.png', 'blue')
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:795: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linewidth"].fillna(1, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:796: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["facecolor"].fillna(self._facecolor, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:797: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["edgecolor"].fillna(styles["facecolor"], inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:798: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linestyle"].fillna("solid", inplace=True)
No description has been provided for this image
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:795: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linewidth"].fillna(1, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:796: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["facecolor"].fillna(self._facecolor, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:797: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["edgecolor"].fillna(styles["facecolor"], inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:798: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linestyle"].fillna("solid", inplace=True)
No description has been provided for this image

Now I use boxem plots and include a line of best fit for the medians

In [7]:
import os
import pandas as pd
import matplotlib.pyplot as plt
from upsetplot import UpSet, from_indicators
from sklearn.linear_model import LinearRegression
import numpy as np

# Function to create the combined UpSet and Violin plot
def create_upset_violin(data, title, output_file, color):
    indicator_cols = ['BM', 'Spleen', 'Ovary', 'Liver', 'Lung', 'Kidney']
    # Ensure all indicator columns are boolean type
    data[indicator_cols] = data[indicator_cols].astype(bool)
    # Filter out rows where ARE_score is NaN and Gene.biotype is not 'protein_coding'
    data = data[data['gene_biotype'] == 'protein_coding']
    data = data.dropna(subset=['ARE_score'])
    data['ARE_score'] = pd.to_numeric(data['ARE_score'], errors='coerce')
    
    # Group by gene ID and aggregate the Boolean indicators
    data_agg = data.groupby('ID')[indicator_cols + ['ARE_score']].max()
    
    # Ensure every row is treated as unique for UpSet plot
    data_agg = data_agg.reset_index()

    # Create the UpSet plot data
    upset_data = from_indicators(indicator_cols, data_agg)
    
    # Initialize UpSet plot with desired formatting
    upset = UpSet(
        upset_data,
        subset_size='count',
        sort_by='-degree',
        facecolor=color,
        show_counts=True,
        # max_subset_size=30 # Limiting the large number of insignificant intersections
    )
    
    # Add the violin plots for ARE_score values
    upset.add_catplot(
        value='ARE_score',
        kind='boxen',
        elements=10,
        color=color
    )

    # Plot UpSet with catplots
    fig = plt.figure(figsize=(10, 6))
    upset.plot(fig=fig)
    
    # Add title for the combined plot
    plt.suptitle(f'{title} Genes with ARE Score Distribution')

    # Calculate mean ARE_score for each subset
    subset_means = upset_data.groupby(upset_data.index)['ARE_score'].mean().reset_index()
    subset_means.columns = ['Subset', 'Mean_ARE_score']
    
    # Prepare data for linear regression
    X = np.arange(len(subset_means)).reshape(-1, 1)
    y = subset_means['Mean_ARE_score'].values
    
    # Perform linear regression
    model = LinearRegression()
    model.fit(X, y)
    y_pred = model.predict(X)
    
    # Plot the line of best fit
    plt.plot(X, y_pred, color='blue', lw=2, linestyle='--')
    
    # Save the combined plot
    plt.savefig(output_file)
    plt.show()

# Load and concatenate data from all updated files in the specified directory
def load_and_concat_data(directory):
    files = [os.path.join(directory, file) for file in os.listdir(directory) if file.endswith('_annotated_updated.csv')]
    data = pd.concat((pd.read_csv(file) for file in files), ignore_index=True)
    return data

# Load upregulated and downregulated data
up_data = load_and_concat_data("upregulated_genes_updated")
down_data = load_and_concat_data("downregulated_genes_updated")

# Create UpSet plots with violin plots and lines of best fit
create_upset_violin(up_data, 'Upregulated', 'upregulated_genes_upset_violin.png', 'red')
create_upset_violin(down_data, 'Downregulated', 'downregulated_genes_upset_violin.png', 'blue')
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:795: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linewidth"].fillna(1, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:796: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["facecolor"].fillna(self._facecolor, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:797: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["edgecolor"].fillna(styles["facecolor"], inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:798: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linestyle"].fillna("solid", inplace=True)
No description has been provided for this image
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:795: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linewidth"].fillna(1, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:796: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["facecolor"].fillna(self._facecolor, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:797: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["edgecolor"].fillna(styles["facecolor"], inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:798: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linestyle"].fillna("solid", inplace=True)
No description has been provided for this image

Same thing just using polynomial line of best fit

In [8]:
import os
import pandas as pd
import matplotlib.pyplot as plt
from upsetplot import UpSet, from_indicators
from sklearn.preprocessing import PolynomialFeatures
from sklearn.linear_model import LinearRegression
import numpy as np

# Function to create the combined UpSet and Violin plot
def create_upset_violin(data, title, output_file, color):
    indicator_cols = ['BM', 'Spleen', 'Ovary', 'Liver', 'Lung', 'Kidney']
    # Ensure all indicator columns are boolean type
    data[indicator_cols] = data[indicator_cols].astype(bool)
    # Filter out rows where ARE_score is NaN and Gene.biotype is not 'protein_coding'
    data = data[data['gene_biotype'] == 'protein_coding']
    data = data.dropna(subset=['ARE_score'])
    data['ARE_score'] = pd.to_numeric(data['ARE_score'], errors='coerce')
    
    # Group by gene ID and aggregate the Boolean indicators
    data_agg = data.groupby('ID')[indicator_cols + ['ARE_score']].max()
    
    # Ensure every row is treated as unique for UpSet plot
    data_agg = data_agg.reset_index()

    # Create the UpSet plot data
    upset_data = from_indicators(indicator_cols, data_agg)
    
    # Initialize UpSet plot with desired formatting
    upset = UpSet(
        upset_data,
        subset_size='count',
        sort_by='-degree',
        facecolor=color,
        show_counts=True,
        # max_subset_size=30 # Limiting the large number of insignificant intersections
    )
    
    # Add the violin plots for ARE_score values
    upset.add_catplot(
        value='ARE_score',
        kind='boxen',
        elements=10,
        color=color
    )

    # Plot UpSet with catplots
    fig = plt.figure(figsize=(10, 6))
    upset.plot(fig=fig)
    
    # Add title for the combined plot
    plt.suptitle(f'{title} Genes with ARE Score Distribution')

    # Calculate mean ARE_score for each subset
    subset_means = upset_data.groupby(upset_data.index)['ARE_score'].mean().reset_index()
    subset_means.columns = ['Subset', 'Mean_ARE_score']
    
    # Prepare data for polynomial regression
    X = np.arange(len(subset_means)).reshape(-1, 1)
    y = subset_means['Mean_ARE_score'].values
    
    # Perform polynomial regression (degree 2 or 3 for better accuracy)
    polynomial_features = PolynomialFeatures(degree=2)
    X_poly = polynomial_features.fit_transform(X)
    model = LinearRegression()
    model.fit(X_poly, y)
    y_poly_pred = model.predict(X_poly)
    
    # Plot the curve of best fit
    plt.plot(X, y_poly_pred, color='blue', lw=2, linestyle='--')
    
    # Save the combined plot
    plt.savefig(output_file)
    plt.show()

# Load and concatenate data from all updated files in the specified directory
def load_and_concat_data(directory):
    files = [os.path.join(directory, file) for file in os.listdir(directory) if file.endswith('_annotated_updated.csv')]
    data = pd.concat((pd.read_csv(file) for file in files), ignore_index=True)
    return data

# Load upregulated and downregulated data
up_data = load_and_concat_data("upregulated_genes_updated")
down_data = load_and_concat_data("downregulated_genes_updated")

# Create UpSet plots with violin plots and polynomial lines of best fit
create_upset_violin(up_data, 'Upregulated', 'upregulated_genes_upset_violin.png', 'red')
create_upset_violin(down_data, 'Downregulated', 'downregulated_genes_upset_violin.png', 'blue')
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:795: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linewidth"].fillna(1, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:796: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["facecolor"].fillna(self._facecolor, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:797: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["edgecolor"].fillna(styles["facecolor"], inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:798: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linestyle"].fillna("solid", inplace=True)
No description has been provided for this image
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:795: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linewidth"].fillna(1, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:796: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["facecolor"].fillna(self._facecolor, inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:797: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["edgecolor"].fillna(styles["facecolor"], inplace=True)
/home/gsgeorge/.local/lib/python3.12/site-packages/upsetplot/plotting.py:798: FutureWarning: A value is trying to be set on a copy of a DataFrame or Series through chained assignment using an inplace method.
The behavior will change in pandas 3.0. This inplace method will never work because the intermediate object on which we are setting values always behaves as a copy.

For example, when doing 'df[col].method(value, inplace=True)', try using 'df.method({col: value}, inplace=True)' or df[col] = df[col].method(value) instead, to perform the operation inplace on the original object.


  styles["linestyle"].fillna("solid", inplace=True)
No description has been provided for this image
In [9]:
import pandas as pd
import seaborn as sns
import matplotlib.pyplot as plt
import glob

# Function to read all CSV files from updated folders
def read_data(regulation_type):
    path = f'{regulation_type}_genes_updated/*.csv'
    filenames = glob.glob(path)

    dfs = []
    for filename in filenames:
        df = pd.read_csv(filename)
        tissue = filename.split('_')[0].split('/')[-1]  # Extract tissue info from the file name
        df['Tissue'] = tissue
        df['Regulation'] = regulation_type.capitalize()
        dfs.append(df)
    
    return pd.concat(dfs, ignore_index=True)

# Reading the data
upregulated_data = read_data("upregulated")
downregulated_data = read_data("downregulated")
unregulated_data = read_data("unregulated")

# Combine all data
combined_data = pd.concat([upregulated_data, downregulated_data, unregulated_data], ignore_index=True)

# Function to calculate the number of intersections and mean ARE score
def prepare_data(df):
    df['NumTissues'] = df[['BM', 'Spleen', 'Ovary', 'Liver', 'Lung', 'Kidney']].sum(axis=1)
    return df[['NumTissues', 'ARE_score', 'Regulation']]

# Preparing data for plotting
prepared_data = prepare_data(combined_data)

# Plotting function
def plot_jitter_violins(data, regulation_type, color):
    plt.figure(figsize=(10, 6))
    sns.violinplot(x='NumTissues', y='ARE_score', data=data[data['Regulation'] == regulation_type.capitalize()], 
                   inner=None, color=color, scale='width', alpha=0.6)
    sns.stripplot(x='NumTissues', y='ARE_score', data=data[data['Regulation'] == regulation_type.capitalize()],
                  jitter=True, color=color, size=2.5)
    plt.xlabel("Number of Tissues")
    plt.ylabel("ARE Score")
    plt.title(f"ARE Score Distribution for {regulation_type.capitalize()} Genes")
    plt.grid(True)
    plt.show()

# Plotting the scores
plot_jitter_violins(prepared_data, "upregulated", "orange")
plot_jitter_violins(prepared_data, "downregulated", "blue")
plot_jitter_violins(prepared_data, "unregulated", "black")
/tmp/ipykernel_2823251/3320125982.py:40: FutureWarning: 

The `scale` parameter has been renamed and will be removed in v0.15.0. Pass `density_norm='width'` for the same effect.
  sns.violinplot(x='NumTissues', y='ARE_score', data=data[data['Regulation'] == regulation_type.capitalize()],
No description has been provided for this image
/tmp/ipykernel_2823251/3320125982.py:40: FutureWarning: 

The `scale` parameter has been renamed and will be removed in v0.15.0. Pass `density_norm='width'` for the same effect.
  sns.violinplot(x='NumTissues', y='ARE_score', data=data[data['Regulation'] == regulation_type.capitalize()],
No description has been provided for this image
/tmp/ipykernel_2823251/3320125982.py:40: FutureWarning: 

The `scale` parameter has been renamed and will be removed in v0.15.0. Pass `density_norm='width'` for the same effect.
  sns.violinplot(x='NumTissues', y='ARE_score', data=data[data['Regulation'] == regulation_type.capitalize()],
No description has been provided for this image
In [10]:
import pandas as pd
import seaborn as sns
import matplotlib.pyplot as plt
import glob

# Function to read all CSV files from updated folders
def read_data(regulation_type):
    path = f'{regulation_type}_genes_updated/*.csv'
    filenames = glob.glob(path)

    dfs = []
    for filename in filenames:
        df = pd.read_csv(filename)
        tissue = filename.split('_')[0].split('/')[-1]  # Extract tissue info from the file name
        df['Tissue'] = tissue
        df['Regulation'] = regulation_type.capitalize()
        dfs.append(df)
    
    return pd.concat(dfs, ignore_index=True)

# Reading the data
upregulated_data = read_data("upregulated")
downregulated_data = read_data("downregulated")
unregulated_data = read_data("unregulated")

# Combine all data
combined_data = pd.concat([upregulated_data, downregulated_data, unregulated_data], ignore_index=True)

# Function to calculate the number of intersections and prepare data for plotting
def prepare_data(df):
    df['NumTissues'] = df[['BM', 'Spleen', 'Ovary', 'Liver', 'Lung', 'Kidney']].sum(axis=1)
    return df[['NumTissues', 'ARE_score', 'Regulation']]

# Preparing data for plotting
prepared_data = prepare_data(combined_data)

# Plotting function
def plot_jitter_violins(ax, data, regulation_type, color):
    sns.violinplot(ax=ax, x='NumTissues', y='ARE_score', data=data[data['Regulation'] == regulation_type],
                   inner=None, color=color, scale='width', alpha=0.6)
    sns.stripplot(ax=ax, x='NumTissues', y='ARE_score', data=data[data['Regulation'] == regulation_type],
                  jitter=True, color=color, size=2.5)
    ax.set_title(f"{regulation_type} Genes", fontsize=14, weight='bold')
    ax.set_xlabel("Number of Tissues", fontsize=12)
    ax.set_ylabel("ARE Score", fontsize=12)

# Creating the figure and subplots
fig, axes = plt.subplots(nrows=3, ncols=1, figsize=(10, 18), sharex=True)
fig.suptitle('ARE Score Distribution Across Number of Tissues', fontsize=16, weight='bold')

# Plotting the scores for each regulation type
plot_jitter_violins(axes[0], prepared_data, "Upregulated", "orange")
plot_jitter_violins(axes[1], prepared_data, "Downregulated", "blue")
plot_jitter_violins(axes[2], prepared_data, "Unregulated", "black")

# Adjust layout
plt.tight_layout(rect=[0, 0, 1, 0.95])  # Adjust subplots to fit title
plt.subplots_adjust(hspace=0.4)

# Adding legends to individual plots
axes[0].legend(['Upregulated'], loc='upper right')
axes[1].legend(['Downregulated'], loc='upper right')
axes[2].legend(['Unregulated'], loc='upper right')

plt.show()
/tmp/ipykernel_2823251/2060466322.py:39: FutureWarning: 

The `scale` parameter has been renamed and will be removed in v0.15.0. Pass `density_norm='width'` for the same effect.
  sns.violinplot(ax=ax, x='NumTissues', y='ARE_score', data=data[data['Regulation'] == regulation_type],
/tmp/ipykernel_2823251/2060466322.py:39: FutureWarning: 

The `scale` parameter has been renamed and will be removed in v0.15.0. Pass `density_norm='width'` for the same effect.
  sns.violinplot(ax=ax, x='NumTissues', y='ARE_score', data=data[data['Regulation'] == regulation_type],
/tmp/ipykernel_2823251/2060466322.py:39: FutureWarning: 

The `scale` parameter has been renamed and will be removed in v0.15.0. Pass `density_norm='width'` for the same effect.
  sns.violinplot(ax=ax, x='NumTissues', y='ARE_score', data=data[data['Regulation'] == regulation_type],
No description has been provided for this image

Corrected tissue specific and overall violin plots

In [11]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import os
from scipy.stats import ks_2samp
from statsmodels.stats.multitest import multipletests

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_annotated_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0] # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and filter for valid ARE scores
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Checking for any gene appearing as both upregulated and downregulated in the same tissue
conflicting_genes = combined_data.groupby(['ID', 'Tissue'])['Regulation'].nunique()
conflicts = conflicting_genes[conflicting_genes > 1]
print(conflicts) # This should be empty if the data is clean and the duplicates are correctly removed

# Conducting a KS-test between the scores of upregulated and downregulated genes
up_scores = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score']
down_scores = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score']
ks_stat, p_value = ks_2samp(up_scores, down_scores)

# Function to add p-value annotations
def add_p_val_annotation(ax, x1, x2, y, p_val, h=0.02):
    p_text = f'p = {p_val:.2e}'
    ax.plot([x1, x1, x2, x2], [y, y + h, y + h, y], color='k')
    ax.text((x1 + x2) * .5, y + h, p_text, ha='center', va='bottom', color='k')

# Function to add mean annotations
def add_mean_annotation(ax, x_pos, mean, color):
    mean_text = f'mean = {mean:.3f}'
    ax.text(x_pos, mean + 0.5, mean_text, ha='center', va='bottom', color=color, fontsize=10, fontweight='bold')

# Set the Seaborn context
sns.set_context("notebook", font_scale=1.25)

# Create a dummy column
combined_data['dummy'] = 'All' # Add a dummy column for overall plotting

# Calculate median ARE scores for upregulated and downregulated genes for each tissue
median_scores = combined_data.groupby(['Tissue', 'Regulation'])['ARE_score'].median().unstack()
median_scores['Difference'] = median_scores['Upregulated'] - median_scores['Downregulated']

# Sort tissues based on the difference in median ARE scores
sorted_tissues = median_scores.sort_values(by='Difference', ascending=False).index

# Create ordered categorical type to sort tissues in the plot
combined_data['Tissue'] = pd.Categorical(combined_data['Tissue'], categories=sorted_tissues, ordered=True)

# Create a figure with two subplots
fig, axes = plt.subplots(nrows=1, ncols=2, figsize=(20, 8))

# Plot for overall ARE score violin plot comparing up and downregulated genes across all tissues
sns.violinplot(data=combined_data, x='dummy', y='ARE_score', hue='Regulation', scale='area',
               palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0])
sns.stripplot(data=combined_data, x='dummy', y='ARE_score', hue='Regulation', dodge=True, jitter=True, alpha=0.6,
              palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0])
axes[0].set_title('ARE Scores Across Tissues by Regulation Type')
axes[0].set_ylabel('ARE Score')
axes[0].set_xlabel("Regulation Type")
axes[0].set_xticklabels(["All Regulation Types"])

# Add means annotation for overall plot
overall_up_mean = up_scores.mean()
overall_down_mean = down_scores.mean()
add_mean_annotation(axes[0], -0.2, overall_up_mean, 'orange')
add_mean_annotation(axes[0], 0.2, overall_down_mean, 'blue')

# Add p-value annotation for overall plot
max_score = max(combined_data['ARE_score'])
add_p_val_annotation(axes[0], x1=-0.2, x2=0.2, y=max_score, p_val=p_value, h=max_score * 0.1)

# Calculate and store p-values for each tissue
p_values = []
for tissue, group in combined_data.groupby('Tissue'):
    up_scores_tissue = group[group['Regulation'] == 'Upregulated']['ARE_score']
    down_scores_tissue = group[group['Regulation'] == 'Downregulated']['ARE_score']
    if not up_scores_tissue.empty and not down_scores_tissue.empty:
        ks_stat, p_value = ks_2samp(up_scores_tissue, down_scores_tissue)
        p_values.append(p_value)
    else:
        p_values.append(1) # Assign a non-significant p-value for cases with empty groups

# Apply multiple testing correction (e.g., Benjamini-Hochberg)
reject, corrected_p_vals, _, _ = multipletests(p_values, method='fdr_bh')

# Plot for ARE score distributions for each tissue, now ordered
violinplot = sns.violinplot(data=combined_data, x='Tissue', y='ARE_score', hue='Regulation', scale='width',
                            palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1])
sns.stripplot(data=combined_data, x='Tissue', y='ARE_score', hue='Regulation', dodge=True, jitter=True, alpha=0.6,
              palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1])
axes[1].set_title('ARE Scores by Tissue and Regulation Type')
axes[1].set_ylabel('ARE Score')
axes[1].set_xlabel('Tissue')
axes[0].get_legend().remove()

# Move Legend Outside
handles, labels = axes[1].get_legend_handles_labels()
axes[1].legend(handles[:2], labels[:2], loc='upper left', bbox_to_anchor=(1, 1), title='Regulation')

# Get positions for corrected p-values
positions = axes[1].get_xticks()
position_map = {group: pos for group, pos in zip(sorted_tissues, positions)}

# Plot corrected p-values on the second subplot
for (tissue, group), p_value in zip(combined_data.groupby('Tissue'), corrected_p_vals):
    if not group[group['Regulation'] == 'Upregulated']['ARE_score'].empty and not group[group['Regulation'] == 'Downregulated']['ARE_score'].empty:
        up_pos = position_map[tissue] - 0.2 # Adjust depending on position
        down_pos = position_map[tissue] + 0.2 # Adjust depending on position
        max_height = max(group['ARE_score'])
        add_p_val_annotation(axes[1], up_pos, down_pos, max_height, p_value, h=max_height * 0.1)

plt.tight_layout()

# Save the figure as a .svg file
plt.savefig("Overall_ARE_distribution_including_zeros.svg", format="svg")
plt.show()
Series([], Name: Regulation, dtype: int64)
/tmp/ipykernel_2823251/2819809908.py:77: FutureWarning: 

The `scale` parameter has been renamed and will be removed in v0.15.0. Pass `density_norm='area'` for the same effect.
  sns.violinplot(data=combined_data, x='dummy', y='ARE_score', hue='Regulation', scale='area',
/tmp/ipykernel_2823251/2819809908.py:84: UserWarning: set_ticklabels() should only be used with a fixed number of ticks, i.e. after set_ticks() or using a FixedLocator.
  axes[0].set_xticklabels(["All Regulation Types"])
/tmp/ipykernel_2823251/2819809908.py:98: FutureWarning: The default of observed=False is deprecated and will be changed to True in a future version of pandas. Pass observed=False to retain current behavior or observed=True to adopt the future default and silence this warning.
  for tissue, group in combined_data.groupby('Tissue'):
/tmp/ipykernel_2823251/2819809908.py:111: FutureWarning: 

The `scale` parameter has been renamed and will be removed in v0.15.0. Pass `density_norm='width'` for the same effect.
  violinplot = sns.violinplot(data=combined_data, x='Tissue', y='ARE_score', hue='Regulation', scale='width',
/tmp/ipykernel_2823251/2819809908.py:129: FutureWarning: The default of observed=False is deprecated and will be changed to True in a future version of pandas. Pass observed=False to retain current behavior or observed=True to adopt the future default and silence this warning.
  for (tissue, group), p_value in zip(combined_data.groupby('Tissue'), corrected_p_vals):
No description has been provided for this image
In [12]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import os
from scipy.stats import ks_2samp
from statsmodels.stats.multitest import multipletests

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_annotated_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0] # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and filter for valid ARE scores
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]
print(min(combined_data))
# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Checking for any gene appearing as both upregulated and downregulated in the same tissue
conflicting_genes = combined_data.groupby(['ID', 'Tissue'])['Regulation'].nunique()
conflicts = conflicting_genes[conflicting_genes > 1]
print(conflicts) # This should be empty if the data is clean and the duplicates are correctly removed

# Conducting a KS-test between the scores of upregulated and downregulated genes
up_scores = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score']
down_scores = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score']
ks_stat, p_value = ks_2samp(up_scores, down_scores)

# Function to add p-value annotations
def add_p_val_annotation(ax, x1, x2, y, p_val, h=0.02):
    p_text = f'p = {p_val:.2e}'
    ax.plot([x1, x1, x2, x2], [y, y + h, y + h, y], color='k')
    ax.text((x1 + x2) * .5, y + h, p_text, ha='center', va='bottom', color='k')

# Function to add mean annotations
def add_mean_annotation(ax, x_pos, mean, color):
    mean_text = f'mean = {mean:.3f}'
    ax.text(x_pos, mean + 0.5, mean_text, ha='center', va='bottom', color=color, fontsize=10, fontweight='bold')

# Set the Seaborn context
sns.set_context("notebook", font_scale=1.25)

# Create a dummy column
combined_data['dummy'] = 'All' # Add a dummy column for overall plotting

# Calculate median ARE scores for upregulated and downregulated genes for each tissue
median_scores = combined_data.groupby(['Tissue', 'Regulation'])['ARE_score'].median().unstack()
median_scores['Difference'] = median_scores['Upregulated'] - median_scores['Downregulated']

# Sort tissues based on the difference in median ARE scores
sorted_tissues = median_scores.sort_values(by='Difference', ascending=False).index

# Create ordered categorical type to sort tissues in the plot
combined_data['Tissue'] = pd.Categorical(combined_data['Tissue'], categories=sorted_tissues, ordered=True)

# Create a figure with two subplots
fig, axes = plt.subplots(nrows=1, ncols=2, figsize=(20, 8))

# Plot for overall ARE score box plot comparing up and downregulated genes across all tissues
sns.boxplot(data=combined_data, x='dummy', y='ARE_score', hue='Regulation', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0], showfliers=False)
sns.stripplot(data=combined_data, x='dummy', y='ARE_score', hue='Regulation', dodge=True, jitter=True, alpha=0.6, palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0])
axes[0].set_title('ARE Scores Across Tissues by Regulation Type')
axes[0].set_ylabel('ARE Score')
axes[0].set_xlabel("Regulation Type")
axes[0].set_xticklabels(["All Regulation Types"])

# Add means annotation for overall plot
overall_up_mean = up_scores.mean()
overall_down_mean = down_scores.mean()
add_mean_annotation(axes[0], -0.2, overall_up_mean, 'orange')
add_mean_annotation(axes[0], 0.2, overall_down_mean, 'blue')

# Add p-value annotation for overall plot
max_score = max(combined_data['ARE_score'])
add_p_val_annotation(axes[0], x1=-0.2, x2=0.2, y=max_score, p_val=p_value, h=max_score * 0.1)

# Calculate and store p-values for each tissue
p_values = []
for tissue, group in combined_data.groupby('Tissue'):
    up_scores_tissue = group[group['Regulation'] == 'Upregulated']['ARE_score']
    down_scores_tissue = group[group['Regulation'] == 'Downregulated']['ARE_score']
    if not up_scores_tissue.empty and not down_scores_tissue.empty:
        ks_stat, p_value = ks_2samp(up_scores_tissue, down_scores_tissue)
        p_values.append(p_value)
    else:
        p_values.append(1) # Assign a non-significant p-value for cases with empty groups

# Apply multiple testing correction (e.g., Benjamini-Hochberg)
reject, corrected_p_vals, _, _ = multipletests(p_values, method='fdr_bh')

# Plot for ARE score distributions for each tissue, now ordered
sns.boxplot(data=combined_data, x='Tissue', y='ARE_score', hue='Regulation', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1], showfliers=False)
sns.stripplot(data=combined_data, x='Tissue', y='ARE_score', hue='Regulation', dodge=True, jitter=True, alpha=0.6, palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1])
axes[1].set_title('ARE Scores by Tissue and Regulation Type')
axes[1].set_ylabel('ARE Score')
axes[1].set_xlabel('Tissue')
axes[0].get_legend().remove()

# Move Legend Outside
handles, labels = axes[1].get_legend_handles_labels()
axes[1].legend(handles[:2], labels[:2], loc='upper left', bbox_to_anchor=(1, 1), title='Regulation')

# Get positions for corrected p-values
positions = axes[1].get_xticks()
position_map = {group: pos for group, pos in zip(sorted_tissues, positions)}

# Plot corrected p-values on the second subplot
for (tissue, group), p_value in zip(combined_data.groupby('Tissue'), corrected_p_vals):
    if not group[group['Regulation'] == 'Upregulated']['ARE_score'].empty and not group[group['Regulation'] == 'Downregulated']['ARE_score'].empty:
        up_pos = position_map[tissue] - 0.2 # Adjust depending on position
        down_pos = position_map[tissue] + 0.2 # Adjust depending on position
        max_height = max(group['ARE_score'])
        add_p_val_annotation(axes[1], up_pos, down_pos, max_height, p_value, h=max_height * 0.1)

plt.tight_layout()

# Save the figure as a .svg file
plt.savefig("Overall_ARE_distribution_including_zeros.svg", format="svg")
plt.show()
5mer_count
Series([], Name: Regulation, dtype: int64)
/tmp/ipykernel_2823251/3324192236.py:82: UserWarning: set_ticklabels() should only be used with a fixed number of ticks, i.e. after set_ticks() or using a FixedLocator.
  axes[0].set_xticklabels(["All Regulation Types"])
/tmp/ipykernel_2823251/3324192236.py:96: FutureWarning: The default of observed=False is deprecated and will be changed to True in a future version of pandas. Pass observed=False to retain current behavior or observed=True to adopt the future default and silence this warning.
  for tissue, group in combined_data.groupby('Tissue'):
/tmp/ipykernel_2823251/3324192236.py:125: FutureWarning: The default of observed=False is deprecated and will be changed to True in a future version of pandas. Pass observed=False to retain current behavior or observed=True to adopt the future default and silence this warning.
  for (tissue, group), p_value in zip(combined_data.groupby('Tissue'), corrected_p_vals):
No description has been provided for this image

Same plot as above just with a sqrt() transformed y-axis

In [13]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import os
from scipy.stats import ks_2samp
from statsmodels.stats.multitest import multipletests
import numpy as np
from matplotlib.ticker import ScalarFormatter

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_annotated_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and filter for valid ARE scores
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Checking for any gene appearing as both upregulated and downregulated in the same tissue
conflicting_genes = combined_data.groupby(['ID', 'Tissue'])['Regulation'].nunique()
conflicts = conflicting_genes[conflicting_genes > 1]
print(conflicts)  # This should be empty if the data is clean and the duplicates are correctly removed

# Conducting a KS-test between the scores of upregulated and downregulated genes
up_scores = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score']
down_scores = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score']
ks_stat, p_value = ks_2samp(up_scores, down_scores)

# Function to add p-value annotations
def add_p_val_annotation(ax, x1, x2, y, p_val, h=0.02):
    p_text = f'p = {p_val:.2e}'
    ax.plot([x1, x1, x2, x2], [y, y + h, y + h, y], color='k')
    ax.text((x1 + x2) * .5, y + h, p_text, ha='center', va='bottom', color='k')

# Function to add mean annotations
def add_mean_annotation(ax, x_pos, mean, color):
    mean_text = f'mean = {mean:.3f}'
    ax.text(x_pos, mean + 0.5, mean_text, ha='center', va='bottom', color=color, fontsize=10, fontweight='bold')

# Set the Seaborn context
sns.set_context("notebook", font_scale=1.25)

# Create a dummy column
combined_data['dummy'] = 'All'  # Add a dummy column for overall plotting

# Apply square root transformation
combined_data['ARE_score_transformed'] = np.sqrt(combined_data['ARE_score'])

# Calculate median ARE scores for upregulated and downregulated genes for each tissue
median_scores = combined_data.groupby(['Tissue', 'Regulation'])['ARE_score_transformed'].median().unstack()
median_scores['Difference'] = median_scores['Upregulated'] - median_scores['Downregulated']

# Sort tissues based on the difference in median ARE scores
sorted_tissues = median_scores.sort_values(by='Difference', ascending=False).index

# Create ordered categorical type to sort tissues in the plot
combined_data['Tissue'] = pd.Categorical(combined_data['Tissue'], categories=sorted_tissues, ordered=True)

# Create a figure with two subplots
fig, axes = plt.subplots(nrows=1, ncols=2, figsize=(20, 8))

# Plot for overall ARE score box plot comparing up and downregulated genes across all tissues
sns.boxplot(data=combined_data, x='dummy', y='ARE_score_transformed', hue='Regulation', 
            palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0], showfliers=False)
sns.stripplot(data=combined_data, x='dummy', y='ARE_score_transformed', hue='Regulation', 
              dodge=True, jitter=True, alpha=0.6, palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0])
axes[0].set_title('ARE Scores Across Tissues by Regulation Type')
axes[0].set_ylabel('ARE Score (Square Root Transformed)')
axes[0].set_xlabel("Regulation Type")
axes[0].set_xticklabels(["All Regulation Types"])

# Add means annotation for overall plot
overall_up_mean = np.sqrt(up_scores).mean()
overall_down_mean = np.sqrt(down_scores).mean()
add_mean_annotation(axes[0], -0.2, overall_up_mean, 'orange')
add_mean_annotation(axes[0], 0.2, overall_down_mean, 'blue')

# Add p-value annotation for overall plot
max_score = max(combined_data['ARE_score_transformed'])
add_p_val_annotation(axes[0], x1=-0.2, x2=0.2, y=max_score, p_val=p_value, h=max_score * 0.1)

# Calculate and store p-values for each tissue
p_values = []
for tissue, group in combined_data.groupby('Tissue'):
    up_scores_tissue = group[group['Regulation'] == 'Upregulated']['ARE_score_transformed']
    down_scores_tissue = group[group['Regulation'] == 'Downregulated']['ARE_score_transformed']
    if not up_scores_tissue.empty and not down_scores_tissue.empty:
        ks_stat, p_value = ks_2samp(up_scores_tissue, down_scores_tissue)
        p_values.append(p_value)
    else:
        p_values.append(1) # Assign a non-significant p-value for cases with empty groups

# Apply multiple testing correction (e.g., Benjamini-Hochberg)
reject, corrected_p_vals, _, _ = multipletests(p_values, method='fdr_bh')

# Plot for ARE score distributions for each tissue, now ordered
sns.boxplot(data=combined_data, x='Tissue', y='ARE_score_transformed', hue='Regulation', 
            palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1], showfliers=False)
sns.stripplot(data=combined_data, x='Tissue', y='ARE_score_transformed', hue='Regulation', 
              dodge=True, jitter=True, alpha=0.6, palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1])
axes[1].set_title('ARE Scores by Tissue and Regulation Type')
axes[1].set_ylabel('ARE Score (Square Root Transformed)')
axes[1].set_xlabel('Tissue')
axes[0].get_legend().remove()

# Move Legend Outside
handles, labels = axes[1].get_legend_handles_labels()
axes[1].legend(handles[:2], labels[:2], loc='upper left', bbox_to_anchor=(1, 1), title='Regulation')

# Get positions for corrected p-values
positions = axes[1].get_xticks()
position_map = {group: pos for group, pos in zip(sorted_tissues, positions)}

# Plot corrected p-values on the second subplot
for (tissue, group), p_value in zip(combined_data.groupby('Tissue'), corrected_p_vals):
    if not group[group['Regulation'] == 'Upregulated']['ARE_score_transformed'].empty and not group[group['Regulation'] == 'Downregulated']['ARE_score_transformed'].empty:
        up_pos = position_map[tissue] - 0.2  # Adjust depending on position
        down_pos = position_map[tissue] + 0.2  # Adjust depending on position
        max_height = max(group['ARE_score_transformed'])
        add_p_val_annotation(axes[1], up_pos, down_pos, max_height, p_value, h=max_height * 0.1)

plt.tight_layout()
# Save the figure as a .svg file
plt.savefig("Overall_ARE_distribution_square_root_transformed.svg", format="svg")
plt.show()
Series([], Name: Regulation, dtype: int64)
/tmp/ipykernel_2823251/777753550.py:89: UserWarning: set_ticklabels() should only be used with a fixed number of ticks, i.e. after set_ticks() or using a FixedLocator.
  axes[0].set_xticklabels(["All Regulation Types"])
/tmp/ipykernel_2823251/777753550.py:103: FutureWarning: The default of observed=False is deprecated and will be changed to True in a future version of pandas. Pass observed=False to retain current behavior or observed=True to adopt the future default and silence this warning.
  for tissue, group in combined_data.groupby('Tissue'):
/tmp/ipykernel_2823251/777753550.py:134: FutureWarning: The default of observed=False is deprecated and will be changed to True in a future version of pandas. Pass observed=False to retain current behavior or observed=True to adopt the future default and silence this warning.
  for (tissue, group), p_value in zip(combined_data.groupby('Tissue'), corrected_p_vals):
No description has been provided for this image

Sqrt transformed:

In [3]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os
from scipy.stats import ks_2samp
from statsmodels.stats.multitest import multipletests

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_annotated_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and filter for valid ARE scores
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Checking for any gene appearing as both upregulated and downregulated in the same tissue
conflicting_genes = combined_data.groupby(['ID', 'Tissue'])['Regulation'].nunique()
conflicts = conflicting_genes[conflicting_genes > 1]
print(conflicts)  # This should be empty if the data is clean and the duplicates are correctly removed

# Conducting a KS-test between the scores of upregulated and downregulated genes
up_scores = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score']
down_scores = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score']
ks_stat, p_value = ks_2samp(up_scores, down_scores)

# Function to add p-value annotations
def add_p_val_annotation(ax, x1, x2, y, p_val, h=0.02):
    p_text = f'p = {p_val:.2e}'
    ax.plot([x1, x1, x2, x2], [y, y + h, y + h, y], color='k')
    ax.text((x1 + x2) * .5, y + h, p_text, ha='center', va='bottom', color='k')

# Function to add mean annotations
def add_mean_annotation(ax, x_pos, mean, color):
    mean_text = f'mean = {mean:.3f}'
    ax.text(x_pos, mean + 0.5, mean_text, ha='center', va='bottom', color=color, fontsize=10, fontweight='bold')

# Set the Seaborn context
sns.set_context("notebook", font_scale=1.25)

# Create a dummy column
combined_data['dummy'] = 'All'  # Add a dummy column for overall plotting

# Calculate median ARE scores for upregulated and downregulated genes for each tissue
median_scores = combined_data.groupby(['Tissue', 'Regulation'])['ARE_score'].median().unstack()
median_scores['Difference'] = median_scores['Upregulated'] - median_scores['Downregulated']

# Sort tissues based on the difference in median ARE scores
sorted_tissues = median_scores.sort_values(by='Difference', ascending=False).index

# Create ordered categorical type to sort tissues in the plot
combined_data['Tissue'] = pd.Categorical(combined_data['Tissue'], categories=sorted_tissues, ordered=True)

# Create a figure with two subplots
fig, axes = plt.subplots(nrows=1, ncols=2, figsize=(20, 8))

# Apply square root transformation to ARE scores
combined_data['sqrt_ARE_score'] = np.sqrt(combined_data['ARE_score'])

# Plot for overall ARE score box plot comparing up and downregulated genes across all tissues
sns.boxplot(data=combined_data, x='dummy', y='sqrt_ARE_score', hue='Regulation', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0], showfliers=False)
sns.stripplot(data=combined_data, x='dummy', y='sqrt_ARE_score', hue='Regulation', dodge=True, jitter=True, alpha=0.6, palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0])
axes[0].set_title('ARE Scores Across Tissues by Regulation Type')
axes[0].set_ylabel('ARE Score (sqrt-transformed)')
axes[0].set_xlabel("Regulation Type")
axes[0].set_xticklabels(["All Regulation Types"])

# Add means annotation for overall plot
overall_up_mean = np.sqrt(overall_up_mean)
overall_down_mean = np.sqrt(overall_down_mean)
add_mean_annotation(axes[0], -0.2, overall_up_mean, 'orange')
add_mean_annotation(axes[0], 0.2, overall_down_mean, 'blue')

# Add p-value annotation for overall plot
max_score = np.sqrt(max(combined_data['ARE_score']))
add_p_val_annotation(axes[0], x1=-0.2, x2=0.2, y=max_score, p_val=p_value, h=max_score * 0.1)

# Calculate and store p-values for each tissue
p_values = []
for tissue, group in combined_data.groupby('Tissue'):
    up_scores_tissue = np.sqrt(group[group['Regulation'] == 'Upregulated']['ARE_score'])
    down_scores_tissue = np.sqrt(group[group['Regulation'] == 'Downregulated']['ARE_score'])
    if not up_scores_tissue.empty and not down_scores_tissue.empty:
        ks_stat, p_value = ks_2samp(up_scores_tissue, down_scores_tissue)
        p_values.append(p_value)
    else:
        p_values.append(1)  # Assign a non-significant p-value for cases with empty groups

# Apply multiple testing correction (e.g., Benjamini-Hochberg)
reject, corrected_p_vals, _, _ = multipletests(p_values, method='fdr_bh')

# Plot for ARE score distributions for each tissue, now ordered
sns.boxplot(data=combined_data, x='Tissue', y='sqrt_ARE_score', hue='Regulation', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1], showfliers=False)
sns.stripplot(data=combined_data, x='Tissue', y='sqrt_ARE_score', hue='Regulation', dodge=True, jitter=True, alpha=0.6, palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1])
axes[1].set_title('ARE Scores by Tissue and Regulation Type')
axes[1].set_ylabel('ARE Score (sqrt-transformed)')
axes[1].set_xlabel('Tissue')
axes[0].get_legend().remove()

# Move Legend Outside
handles, labels = axes[1].get_legend_handles_labels()
axes[1].legend(handles[:2], labels[:2], loc='upper left', bbox_to_anchor=(1, 1), title='Regulation')

# Get positions for corrected p-values
positions = axes[1].get_xticks()
position_map = {group: pos for group, pos in zip(sorted_tissues, positions)}

# Plot corrected p-values on the second subplot
for (tissue, group), p_value in zip(combined_data.groupby('Tissue'), corrected_p_vals):
    if not group[group['Regulation'] == 'Upregulated']['ARE_score'].empty and not group[group['Regulation'] == 'Downregulated']['ARE_score'].empty:
        up_pos = position_map[tissue] - 0.2  # Adjust depending on position
        down_pos = position_map[tissue] + 0.2  # Adjust depending on position
        max_height = max(group['sqrt_ARE_score'])
        add_p_val_annotation(axes[1], up_pos, down_pos, max_height, p_value, h=max_height * 0.1)

plt.tight_layout()

# Save the figure as a .svg file
plt.savefig("/mnt/work_1/gsgeorge/mus_musculus/TPM_analysis/Overall_ARE_distribution_including_zeros.svg", format="svg")
plt.show()
Series([], Name: Regulation, dtype: int64)
/tmp/ipykernel_303325/1986591938.py:86: UserWarning: set_ticklabels() should only be used with a fixed number of ticks, i.e. after set_ticks() or using a FixedLocator.
  axes[0].set_xticklabels(["All Regulation Types"])
/tmp/ipykernel_303325/1986591938.py:100: FutureWarning: The default of observed=False is deprecated and will be changed to True in a future version of pandas. Pass observed=False to retain current behavior or observed=True to adopt the future default and silence this warning.
  for tissue, group in combined_data.groupby('Tissue'):
/tmp/ipykernel_303325/1986591938.py:129: FutureWarning: The default of observed=False is deprecated and will be changed to True in a future version of pandas. Pass observed=False to retain current behavior or observed=True to adopt the future default and silence this warning.
  for (tissue, group), p_value in zip(combined_data.groupby('Tissue'), corrected_p_vals):
No description has been provided for this image

Including 0 in visualization and statistical testing

In [14]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import os
from scipy.stats import ks_2samp
from statsmodels.stats.multitest import multipletests

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_annotated_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and filter for valid ARE scores
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Checking for any gene appearing as both upregulated and downregulated in the same tissue
conflicting_genes = combined_data.groupby(['ID', 'Tissue'])['Regulation'].nunique()
conflicts = conflicting_genes[conflicting_genes > 1]
print(conflicts)  # This should be empty if the data is clean and the duplicates are correctly removed

# Conducting a KS-test between the scores of upregulated and downregulated genes
up_scores = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score']
down_scores = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score']
ks_stat, p_value = ks_2samp(up_scores, down_scores)

# Function to add p-value annotations
def add_p_val_annotation(ax, x1, x2, y, p_val, h=0.02):
    p_text = f'p = {p_val:.2e}'
    ax.plot([x1, x1, x2, x2], [y, y + h, y + h, y], color='k')
    ax.text((x1 + x2) * .5, y + h, p_text, ha='center', va='bottom', color='k')

# Function to add mean annotations
def add_mean_annotation(ax, x_pos, mean, color):
    mean_text = f'mean = {mean:.3f}'
    ax.text(x_pos, mean + 30, mean_text, ha='center', va='bottom', color=color, fontsize=10, fontweight='bold')

# Set the Seaborn context
sns.set_context("notebook", font_scale=1.25)

# Create a dummy column
combined_data['dummy'] = 'All'  # Add a dummy column for overall plotting

# Calculate median ARE scores for upregulated and downregulated genes for each tissue
median_scores = combined_data.groupby(['Tissue', 'Regulation'])['ARE_score'].median().unstack()
median_scores['Difference'] = median_scores['Upregulated'] - median_scores['Downregulated']

# Sort tissues based on the difference in median ARE scores
sorted_tissues = median_scores.sort_values(by='Difference', ascending=False).index

# Create ordered categorical type to sort tissues in the plot
combined_data['Tissue'] = pd.Categorical(combined_data['Tissue'], categories=sorted_tissues, ordered=True)

# Create a figure with two subplots
fig, axes = plt.subplots(nrows=1, ncols=2, figsize=(20, 8))

# Plot for overall ARE score violin plot comparing up and downregulated genes across all tissues
sns.violinplot(data=combined_data, x='dummy', y='ARE_score', hue='Regulation', split=True,
               palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0])
sns.stripplot(data=combined_data, x='dummy', y='ARE_score', hue='Regulation', dodge=True, jitter=True, alpha=0.6,
              palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0])
axes[0].set_title('ARE Scores Across Tissues by Regulation Type')
axes[0].set_ylabel('ARE Score')
axes[0].set_xlabel("Regulation Type")
axes[0].set_xticklabels(["All Upregulated Coding 3' UTR Sequences"])

# Add means annotation for overall plot
overall_up_mean = up_scores.mean()
overall_down_mean = down_scores.mean()
add_mean_annotation(axes[0], -0.2, overall_up_mean, 'orange')
add_mean_annotation(axes[0], 0.2, overall_down_mean, 'blue')

# Add p-value annotation for overall plot
add_p_val_annotation(axes[0], x1=-0.2, x2=0.2, y=max(combined_data['ARE_score']), p_val=p_value, h=max(combined_data['ARE_score']) * 0.1)

# Calculate and store p-values for each tissue
p_values = []
for tissue, group in combined_data.groupby('Tissue', observed=False):
    wt_scores = group[group['Regulation'] == 'Upregulated']['ARE_score']
    ko_scores = group[group['Regulation'] == 'Downregulated']['ARE_score']
    if not wt_scores.empty and not ko_scores.empty:
        ks_stat, p_value = ks_2samp(wt_scores, ko_scores)
        p_values.append(p_value)
    else:
        p_values.append(1)  # Assign a non-significant p-value for cases with empty groups

# Apply multiple testing correction (e.g., Benjamini-Hochberg)
reject, corrected_p_vals, _, _ = multipletests(p_values, method='fdr_bh')

# Plot for ARE score distributions for each tissue, now ordered
violinplot = sns.violinplot(data=combined_data, x='Tissue', y='ARE_score', hue='Regulation', split=True,
                            palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1])
sns.stripplot(data=combined_data, x='Tissue', y='ARE_score', hue='Regulation', dodge=True, jitter=True, alpha=0.6,
              palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1])
axes[1].set_title('ARE Scores by Tissue and Regulation Type')
axes[1].set_ylabel('ARE Score')
axes[1].set_xlabel('Tissue')
axes[0].get_legend().remove()

# Move Legend Outside
handles, labels = axes[1].get_legend_handles_labels()
axes[1].legend(handles[:2], labels[:2], loc='upper left', bbox_to_anchor=(1, 1), title='Regulation')

# Get positions for corrected p-values
positions = violinplot.get_xticks()
position_map = {group: pos for group, pos in zip(sorted_tissues, positions)}

# Plot corrected p-values on the second subplot
for (tissue, group), p_value in zip(combined_data.groupby('Tissue', observed=False), corrected_p_vals):
    if not group[group['Regulation'] == 'Upregulated']['ARE_score'].empty and not group[group['Regulation'] == 'Downregulated']['ARE_score'].empty:
        wt_pos = position_map[tissue] - 0.2  # Adjust depending on position
        ko_pos = position_map[tissue] + 0.2  # Adjust depending on position
        max_height = max(group['ARE_score'])
        add_p_val_annotation(axes[1], wt_pos, ko_pos, max_height, p_value, h=max_height * 0.1)

plt.tight_layout()

# Save the figure as a .svg file
plt.savefig("Overall_ARE_distribution_including_zeros.svg", format="svg")
plt.show()
Series([], Name: Regulation, dtype: int64)
/tmp/ipykernel_2823251/2829530817.py:84: UserWarning: set_ticklabels() should only be used with a fixed number of ticks, i.e. after set_ticks() or using a FixedLocator.
  axes[0].set_xticklabels(["All Upregulated Coding 3' UTR Sequences"])
No description has been provided for this image

Using a mann-whit U test here

In [15]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import os
from scipy.stats import mannwhitneyu
from statsmodels.stats.multitest import multipletests

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_annotated_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and filter for valid ARE scores
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Checking for any gene appearing as both upregulated and downregulated in the same tissue
conflicting_genes = combined_data.groupby(['ID', 'Tissue'])['Regulation'].nunique()
conflicts = conflicting_genes[conflicting_genes > 1]
print(conflicts)  # This should be empty if the data is clean and the duplicates are correctly removed

# Conducting a Mann-Whitney U test between the scores of upregulated and downregulated genes
up_scores = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score']
down_scores = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score']
u_stat, p_value = mannwhitneyu(up_scores, down_scores, alternative='two-sided')

# Function to add p-value annotations
def add_p_val_annotation(ax, x1, x2, y, p_val, h=0.02):
    p_text = f'p = {p_val:.2e}'
    ax.plot([x1, x1, x2, x2], [y, y + h, y + h, y], color='k')
    ax.text((x1 + x2) * .5, y + h, p_text, ha='center', va='bottom', color='k')

# Function to add mean annotations
def add_mean_annotation(ax, x_pos, mean, color):
    mean_text = f'mean = {mean:.3f}'
    ax.text(x_pos, mean + 30, mean_text, ha='center', va='bottom', color=color, fontsize=10, fontweight='bold')

# Set the Seaborn context
sns.set_context("notebook", font_scale=1.25)

# Create a dummy column
combined_data['dummy'] = 'All'  # Add a dummy column for overall plotting

# Calculate median ARE scores for upregulated and downregulated genes for each tissue
median_scores = combined_data.groupby(['Tissue', 'Regulation'])['ARE_score'].median().unstack()
median_scores['Difference'] = median_scores['Upregulated'] - median_scores['Downregulated']

# Sort tissues based on the difference in median ARE scores
sorted_tissues = median_scores.sort_values(by='Difference', ascending=False).index

# Create ordered categorical type to sort tissues in the plot
combined_data['Tissue'] = pd.Categorical(combined_data['Tissue'], categories=sorted_tissues, ordered=True)

# Create a figure with two subplots
fig, axes = plt.subplots(nrows=1, ncols=2, figsize=(20, 8))

# Plot for overall ARE score violin plot comparing up and downregulated genes across all tissues
sns.violinplot(data=combined_data, x='dummy', y='ARE_score', hue='Regulation', split=True,
               palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0])
sns.stripplot(data=combined_data, x='dummy', y='ARE_score', hue='Regulation', dodge=True, jitter=True, alpha=0.6,
              palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0])
axes[0].set_title('ARE Scores Across Tissues by Regulation Type')
axes[0].set_ylabel('ARE Score')
axes[0].set_xlabel("Regulation Type")
axes[0].set_xticklabels(["All Upregulated Coding 3' UTR Sequences"])

# Add means annotation for overall plot
overall_up_mean = up_scores.mean()
overall_down_mean = down_scores.mean()
add_mean_annotation(axes[0], -0.2, overall_up_mean, 'orange')
add_mean_annotation(axes[0], 0.2, overall_down_mean, 'blue')

# Add p-value annotation for overall plot
add_p_val_annotation(axes[0], x1=-0.2, x2=0.2, y=max(combined_data['ARE_score']), p_val=p_value, h=max(combined_data['ARE_score']) * 0.1)

# Calculate and store p-values for each tissue
p_values = []
for tissue, group in combined_data.groupby('Tissue', observed=False):
    wt_scores = group[group['Regulation'] == 'Upregulated']['ARE_score']
    ko_scores = group[group['Regulation'] == 'Downregulated']['ARE_score']
    if not wt_scores.empty and not ko_scores.empty:
        u_stat, p_value = mannwhitneyu(wt_scores, ko_scores, alternative='two-sided')
        p_values.append(p_value)
    else:
        p_values.append(1)  # Assign a non-significant p-value for cases with empty groups

# Apply multiple testing correction (e.g., Benjamini-Hochberg)
reject, corrected_p_vals, _, _ = multipletests(p_values, method='fdr_bh')

# Plot for ARE score distributions for each tissue, now ordered
violinplot = sns.violinplot(data=combined_data, x='Tissue', y='ARE_score', hue='Regulation', split=True,
                            palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1])
sns.stripplot(data=combined_data, x='Tissue', y='ARE_score', hue='Regulation', dodge=True, jitter=True, alpha=0.6,
              palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1])
axes[1].set_title('ARE Scores by Tissue and Regulation Type')
axes[1].set_ylabel('ARE Score')
axes[1].set_xlabel('Tissue')
axes[0].get_legend().remove()

# Move Legend Outside
handles, labels = axes[1].get_legend_handles_labels()
axes[1].legend(handles[:2], labels[:2], loc='upper left', bbox_to_anchor=(1, 1), title='Regulation')

# Get positions for corrected p-values
positions = violinplot.get_xticks()
position_map = {group: pos for group, pos in zip(sorted_tissues, positions)}

# Plot corrected p-values on the second subplot
for (tissue, group), p_value in zip(combined_data.groupby('Tissue', observed=False), corrected_p_vals):
    if not group[group['Regulation'] == 'Upregulated']['ARE_score'].empty and not group[group['Regulation'] == 'Downregulated']['ARE_score'].empty:
        wt_pos = position_map[tissue] - 0.2  # Adjust depending on position
        ko_pos = position_map[tissue] + 0.2  # Adjust depending on position
        max_height = max(group['ARE_score'])
        add_p_val_annotation(axes[1], wt_pos, ko_pos, max_height, p_value, h=max_height * 0.1)

plt.tight_layout()

# Save the figure as a .svg file
plt.savefig("Overall_ARE_distribution_including_zeros.svg", format="svg")
plt.show()
Series([], Name: Regulation, dtype: int64)
/tmp/ipykernel_2823251/3742372327.py:84: UserWarning: set_ticklabels() should only be used with a fixed number of ticks, i.e. after set_ticks() or using a FixedLocator.
  axes[0].set_xticklabels(["All Upregulated Coding 3' UTR Sequences"])
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In [16]:
# Calculate mean ARE scores for upregulated and downregulated genes for each tissue
mean_scores = combined_data.groupby(['Tissue', 'Regulation'])['ARE_score'].mean().unstack()
mean_scores['Difference'] = mean_scores['Upregulated'] - mean_scores['Downregulated']
mean_scores = mean_scores.sort_values(by='Difference', ascending=False)
print(mean_scores)
Regulation  Downregulated  Upregulated  Difference
Tissue                                            
Liver            2.335407     4.030804    1.695397
Kidney           1.720312     3.313000    1.592688
Ovary            1.788636     2.862273    1.073636
BM               3.035843     3.911170    0.875326
Spleen           3.086970     3.665610    0.578640
Lung             3.046296     2.980159   -0.066138
/tmp/ipykernel_2823251/308238176.py:2: FutureWarning: The default of observed=False is deprecated and will be changed to True in a future version of pandas. Pass observed=False to retain current behavior or observed=True to adopt the future default and silence this warning.
  mean_scores = combined_data.groupby(['Tissue', 'Regulation'])['ARE_score'].mean().unstack()
In [17]:
# Calculate mean ARE scores for upregulated and downregulated genes for each tissue
mean_scores = combined_data.groupby(['Tissue', 'Regulation'])['ARE_score'].median().unstack()
mean_scores['Difference'] = mean_scores['Upregulated'] - mean_scores['Downregulated']
mean_scores = mean_scores.sort_values(by='Difference', ascending=False)
print(mean_scores)
Regulation  Downregulated  Upregulated  Difference
Tissue                                            
Kidney                0.0          1.3         1.3
BM                    1.3          2.3         1.0
Lung                  1.3          2.3         1.0
Spleen                1.3          2.0         0.7
Liver                 1.0          1.3         0.3
Ovary                 1.0          1.3         0.3
/tmp/ipykernel_2823251/3645416548.py:2: FutureWarning: The default of observed=False is deprecated and will be changed to True in a future version of pandas. Pass observed=False to retain current behavior or observed=True to adopt the future default and silence this warning.
  mean_scores = combined_data.groupby(['Tissue', 'Regulation'])['ARE_score'].median().unstack()

This is where I realized that the medians would allow me to order the box and whisker plots correctly based on their ARE score medians, which denote the white line down the boxes

Up and downregulated overall pie charts to be added to the overall volcano plot in post using adobe illustrator

In [18]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import os
from scipy.stats import ttest_ind

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Checking for any gene appearing as both upregulated and downregulated in the same tissue
conflicting_genes = combined_data.groupby(['ID', 'Tissue'])['Regulation'].nunique()
conflicts = conflicting_genes[conflicting_genes > 1]
print(conflicts)  # This should be empty if the data is clean and the duplicates are correctly removed

# Proportion for upregulated genes
up_scores = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score']
up_zero_count = (up_scores == 0).sum()
up_nonzero_count = (up_scores > 0).sum()

# Proportion for downregulated genes
down_scores = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score']
down_zero_count = (down_scores == 0).sum()
down_nonzero_count = (down_scores > 0).sum()

# Set the Seaborn context
sns.set_context("notebook", font_scale=1.25)

# Function to plot pie charts and save them
def plot_and_save_pie_chart(labels, sizes, colors, filename):
    fig, ax = plt.subplots()

    def autopct_func(pct, allvals):
        absolute = int(round(pct/100.*sum(allvals)))
        return "{:.1f}%\n({:d})".format(pct, absolute)

    wedges, texts, autotexts = ax.pie(sizes, labels=labels, colors=colors, autopct=lambda pct: autopct_func(pct, sizes), startangle=90, textprops=dict(color="black"))
    
    for text in autotexts:
        # Move the text higher so it fits better
        text.set_position((text.get_position()[0], text.get_position()[1] - 0.2))
    
    ax.axis('equal')  # Equal aspect ratio ensures that pie is drawn as a circle.
    plt.savefig(filename, format='svg')
    plt.show()  # Close the figure to prevent it from displaying immediately and to free memory

# Plot and save upregulated pie chart
up_labels = ['ARE Score = 0', 'ARE Score > 0']
up_sizes = [up_zero_count, up_nonzero_count]
up_colors = ['lightcoral', 'orange']
plot_and_save_pie_chart(up_labels, up_sizes, up_colors, 'upregulated_overall_pie.svg')

# Plot and save downregulated pie chart
down_labels = ['ARE Score = 0', 'ARE Score > 0']
down_sizes = [down_zero_count, down_nonzero_count]
down_colors = ['lightblue', 'blue']
plot_and_save_pie_chart(down_labels, down_sizes, down_colors, 'downregulated_overall_pie.svg')
Series([], Name: Regulation, dtype: int64)
No description has been provided for this image
No description has been provided for this image
In [19]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import os
from scipy.stats import ttest_ind

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0] # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Checking for any gene appearing as both upregulated and downregulated in the same tissue
conflicting_genes = combined_data.groupby(['ID', 'Tissue'])['Regulation'].nunique()
conflicts = conflicting_genes[conflicting_genes > 1]
print(conflicts) # This should be empty if the data is clean and the duplicates are correctly removed

# Categorize the ARE scores into different categories
def categorize_scores(scores):
    categories = pd.cut(scores, bins=[-1, 0, 2, 5, float('inf')], labels=['ARE Score = 0', '0 < ARE Score ≤ 2', '2 < ARE Score < 5', 'ARE Score ≥ 5'])
    return categories.value_counts()

# Proportion for upregulated genesplt.savefig('/mnt/work_1/gsgeorge/mus_musculus/TPM_analysis/TPM_distribution_across_tissues.svg', format='svg')
    def autopct_func(pct, allvals):
        absolute = int(round(pct / 100. * sum(allvals)))
        return "{:.1f}%\n({:d})".format(pct, absolute)
    wedges, texts, autotexts = ax.pie(categories.values, labels=categories.index, colors=colors, autopct=lambda pct: autopct_func(pct, categories.values), startangle=90, textprops=dict(color="black"))
    for text in autotexts:
        # Move the text higher so it fits better
        text.set_position((text.get_position()[0], text.get_position()[1] - 0.2))
    ax.axis('equal') # Equal aspect ratio ensures that pie is drawn as a circle.
    plt.savefig(filename, format='svg')
    plt.show() # Close the figure to prevent it from displaying immediately and to free memory

# Plot and save upregulated pie chart
up_colors = ['lightcoral', 'orange', 'gold', 'darkred']
plot_and_save_pie_chart(up_categories, up_colors, 'upregulated_overall_pie.svg')

# Plot and save downregulated pie chart
down_colors = ['lightblue', 'blue', 'darkblue', 'navy']
plot_and_save_pie_chart(down_categories, down_colors, 'downregulated_overall_pie.svg')
Series([], Name: Regulation, dtype: int64)
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No description has been provided for this image
In [20]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import os
from scipy.stats import ttest_ind

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0] # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Checking for any gene appearing as both upregulated and downregulated in the same tissue
conflicting_genes = combined_data.groupby(['ID', 'Tissue'])['Regulation'].nunique()
conflicts = conflicting_genes[conflicting_genes > 1]
print(conflicts) # This should be empty if the data is clean and the duplicates are correctly removed

# Categorize the ARE scores into different categories
def categorize_scores(scores):
    categories = pd.cut(scores, bins=[-1, 0, 4, float('inf')], labels=['ARE Score = 0', '0 < ARE Score ≤ 4', 'ARE Score > 4'])
    return categories.value_counts()

# Proportion for upregulated genes
up_scores = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score']
up_categories = categorize_scores(up_scores)

# Proportion for downregulated genes
down_scores = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score']
down_categories = categorize_scores(down_scores)

# Set the Seaborn context
sns.set_context("notebook", font_scale=1.25)

# Function to plot pie charts and save them
def plot_and_save_pie_chart(categories, colors, filename, startangle=90, explode=(0, 0, 0.1)):
    fig, ax = plt.subplots()
    def autopct_func(pct, allvals):
        absolute = int(round(pct / 100. * sum(allvals)))
        return "{:.1f}%\n({:d})".format(pct, absolute)
    wedges, texts, autotexts = ax.pie(
        categories.values, 
        labels=categories.index, 
        colors=colors, 
        autopct=lambda pct: autopct_func(pct, categories.values), 
        startangle=startangle, 
        textprops=dict(color="black"),
        explode=explode,
        wedgeprops={"edgecolor": "black", "linewidth": 1}
    )
    for text in autotexts:
        # Move the text higher so it fits better
        text.set_position((text.get_position()[0], text.get_position()[1] - 0.2))
    ax.axis('equal') # Equal aspect ratio ensures that pie is drawn as a circle.
    plt.savefig(filename, format='svg')
    plt.show() # Close the figure to prevent it from displaying immediately and to free memory

# Set a common start angle for consistency
start_angle = 90

# Explode the 'ARE Score = 0' slice larger than the other slices
explode_factor = (0.1, 0, 0)

# Plot and save upregulated pie chart with a gradient from light orange to dark orange
up_colors = ['orange', 'darkorange', 'orangered']
plot_and_save_pie_chart(up_categories, up_colors, 'upregulated_overall_pie.svg', startangle=start_angle, explode=explode_factor)

# Plot and save downregulated pie chart with a gradient from light blue to dark blue
down_colors = ['lightblue', 'blue', 'darkblue']
plot_and_save_pie_chart(down_categories, down_colors, 'downregulated_overall_pie.svg', startangle=start_angle, explode=explode_factor)
Series([], Name: Regulation, dtype: int64)
No description has been provided for this image
No description has been provided for this image
In [21]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import os
from scipy.stats import ttest_ind

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Checking for any gene appearing as both upregulated and downregulated in the same tissue
conflicting_genes = combined_data.groupby(['ID', 'Tissue'])['Regulation'].nunique()
conflicts = conflicting_genes[conflicting_genes > 1]
print(conflicts)  # This should be empty if the data is clean and the duplicates are correctly removed

# Categorize the ARE scores into different categories
def categorize_scores(scores):
    categories = pd.cut(scores, bins=[-1, 0, 2, 5, float('inf')], labels=['ARE Score = 0', '0 < ARE Score ≤ 2', '2 < ARE Score < 5', 'ARE Score ≥ 5'])
    return categories.value_counts()

# Proportion for upregulated genes
up_scores = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score']
up_categories = categorize_scores(up_scores)

# Proportion for downregulated genes
down_scores = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score']
down_categories = categorize_scores(down_scores)

# Set the Seaborn context
sns.set_context("notebook", font_scale=1.5)

# Function to plot pie charts and save them
def plot_and_save_pie_chart(categories, colors, filename):
    fig, ax = plt.subplots()
    def autopct_func(pct, allvals):
        absolute = int(round(pct / 100. * sum(allvals)))
        return "{:.1f}%\n({:d})".format(pct, absolute)
    wedges, texts, autotexts = ax.pie(categories.values, labels=categories.index, colors=colors, 
                                       autopct=lambda pct: autopct_func(pct, categories.values), startangle=90)
    for text in autotexts:
        # Move the text higher so it fits better
        text.set_position((text.get_position()[0], text.get_position()[1] - 0.2))
        text.set_color('white')
    ax.axis('equal')  # Equal aspect ratio ensures that pie is drawn as a circle.
    plt.savefig(filename, format='svg')
    plt.show()  # Close the figure to prevent it from displaying immediately and to free memory

# Plot and save upregulated pie chart
up_colors = ['#FFD700', '#FF4500', '#FF8C00', '#FFA500']  # Golden rod to dark orange
plot_and_save_pie_chart(up_categories, up_colors, 'upregulated_overall_pie.svg')

# Plot and save downregulated pie chart
down_colors = ['#ADD8E6', '#00008B', '#4682B4', '#87CEEB']  # Light blue to dark blue
plot_and_save_pie_chart(down_categories, down_colors, 'downregulated_overall_pie.svg')
Series([], Name: Regulation, dtype: int64)
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No description has been provided for this image

Now I am going to exclude kidney and lung and see what happens:

In [22]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import os
from scipy.stats import ks_2samp

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_annotated_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and filter for valid ARE scores
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Filtering out Lung and Kidney tissues
combined_data = combined_data[~combined_data['Tissue'].isin(['Lung', 'Kidney'])]

# Checking for any gene appearing as both upregulated and downregulated in the same tissue
conflicting_genes = combined_data.groupby(['ID', 'Tissue'])['Regulation'].nunique()
conflicts = conflicting_genes[conflicting_genes > 1]
print(conflicts)  # This should be empty if the data is clean and the duplicates are correctly removed

# Identify the lowest ARE score for upregulated and downregulated sets
lowest_up_score = combined_data[(combined_data['Regulation'] == 'Upregulated') & (combined_data['ARE_score'] > 0)]['ARE_score'].min()
lowest_down_score = combined_data[(combined_data['Regulation'] == 'Downregulated') & (combined_data['ARE_score'] > 0)]['ARE_score'].min()
print(f"Lowest ARE score in upregulated set: {lowest_up_score}")
print(f"Lowest ARE score in downregulated set: {lowest_down_score}")

# Ensure there are no zero scores involved in statistical calculation
assert lowest_up_score > 0, "There are zero scores in the upregulated set."
assert lowest_down_score > 0, "There are zero scores in the downregulated set."

# Conducting a KS-test between the scores of upregulated and downregulated genes
up_scores = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score']
down_scores = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score']
ks_stat, p_value = ks_2samp(up_scores, down_scores)

# Function to add p-value annotations
def add_p_val_annotation(ax, x1, x2, y, p_val, h=0.02):
    p_text = f'p = {p_val:.2e}'
    ax.plot([x1, x1, x2, x2], [y, y + h, y + h, y], color='k')
    ax.text((x1 + x2) * .5, y + h, p_text, ha='center', va='bottom', color='k')

# Function to add mean annotations
def add_mean_annotation(ax, x_pos, mean, color):
    mean_text = f'mean = {mean:.3f}'
    ax.text(x_pos, mean + 30, mean_text, ha='center', va='bottom', color=color, fontsize=10, fontweight='bold')

# Set the Seaborn context
sns.set_context("notebook", font_scale=1)

# Create a dummy column
combined_data['dummy'] = 'All'  # Add a dummy column for overall plotting

# Calculate median ARE scores for upregulated and downregulated genes for each tissue
median_scores = combined_data.groupby(['Tissue', 'Regulation'])['ARE_score'].median().unstack()
median_scores['Difference'] = median_scores['Upregulated'] - median_scores['Downregulated']

# Sort tissues based on the difference in median ARE scores
sorted_tissues = median_scores.sort_values(by='Difference', ascending=False).index

# Create an ordered categorical type to sort tissues in the plot
combined_data['Tissue'] = pd.Categorical(combined_data['Tissue'], categories=sorted_tissues, ordered=True)

# Create a figure with two subplots
fig, axes = plt.subplots(nrows=2, ncols=1, figsize=(8.5, 11))

# Plot for overall ARE score violin plot comparing up and downregulated genes across all tissues
sns.violinplot(data=combined_data, x='dummy', y='ARE_score', hue='Regulation', split=True,
               palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0])
sns.stripplot(data=combined_data, x='dummy', y='ARE_score', hue='Regulation', dodge=True, jitter=True, alpha=0.6,
              palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[0])
axes[0].set_title('ARE Scores Across Tissues by Regulation Type')
axes[0].set_ylabel('ARE Score')
axes[0].set_xlabel("Regulation Type")
axes[0].set_xticklabels(["All Regulated Coding 3' UTR Sequences"])

# Add means annotation for overall plot
overall_up_mean = up_scores.mean()
overall_down_mean = down_scores.mean()
add_mean_annotation(axes[0], -0.2, overall_up_mean, 'orange')
add_mean_annotation(axes[0], 0.2, overall_down_mean, 'blue')

# Add p-value annotation for overall plot
add_p_val_annotation(axes[0], x1=-0.2, x2=0.2, y=max(combined_data['ARE_score']), p_val=p_value, h=max(combined_data['ARE_score']) * 0.1)

# Plot for ARE score distributions for each tissue, now ordered
violinplot = sns.violinplot(data=combined_data, x='Tissue', y='ARE_score', hue='Regulation', split=True,
                            palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1])
sns.stripplot(data=combined_data, x='Tissue', y='ARE_score', hue='Regulation', dodge=True, jitter=True, alpha=0.6,
              palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=axes[1])
axes[1].set_ylabel('ARE Score')
axes[1].set_xlabel('Tissue')
axes[0].get_legend().remove()
axes[1].set_title("ARE Scores by Tissue")

# Move Legend Outside
handles, labels = axes[1].get_legend_handles_labels()
axes[1].legend(handles[:2], labels[:2], loc='upper left', title='Regulation')

# Calculate and plot p-values for each tissue in the second subplot
positions = violinplot.get_xticks()
position_map = {group: pos for group, pos in zip(sorted_tissues, positions)}
grouped = combined_data.groupby('Tissue')
for tissue, group in grouped:
    wt_scores = group[group['Regulation'] == 'Upregulated']['ARE_score']
    ko_scores = group[group['Regulation'] == 'Downregulated']['ARE_score']
    ks_stat, p_value = ks_2samp(wt_scores, ko_scores)
    if not wt_scores.empty and not ko_scores.empty:
        wt_pos = position_map[tissue] - 0.2  # Adjust depending on position
        ko_pos = position_map[tissue] + 0.2  # Adjust depending on position
        max_height = max(group['ARE_score'])
        add_p_val_annotation(axes[1], wt_pos, ko_pos, max_height, p_value, h=max_height * 0.1)

plt.tight_layout()

# Save the figure as a .svg file
plt.savefig("Overall_ARE_distribution.svg", format="svg")
plt.show()
Series([], Name: Regulation, dtype: int64)
Lowest ARE score in upregulated set: 1.0
Lowest ARE score in downregulated set: 1.0
/tmp/ipykernel_2823251/429978461.py:96: UserWarning: set_ticklabels() should only be used with a fixed number of ticks, i.e. after set_ticks() or using a FixedLocator.
  axes[0].set_xticklabels(["All Regulated Coding 3' UTR Sequences"])
/tmp/ipykernel_2823251/429978461.py:124: FutureWarning: The default of observed=False is deprecated and will be changed to True in a future version of pandas. Pass observed=False to retain current behavior or observed=True to adopt the future default and silence this warning.
  grouped = combined_data.groupby('Tissue')
No description has been provided for this image
In [23]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import os
from scipy.stats import ttest_ind

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0] # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Exclude genes found in Lung and Kidney tissues
excluded_tissues = ['Lung', 'Kidney']
combined_data = combined_data[~combined_data['Tissue'].isin(excluded_tissues)]

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Checking for any gene appearing as both upregulated and downregulated in the same tissue
conflicting_genes = combined_data.groupby(['ID', 'Tissue'])['Regulation'].nunique()
conflicts = conflicting_genes[conflicting_genes > 1]
print(conflicts) # This should be empty if the data is clean and the duplicates are correctly removed

# Categorize the ARE scores into different categories
def categorize_scores(scores):
    categories = pd.cut(scores, bins=[-1, 0, 4, float('inf')], labels=['ARE Score = 0', '0 < ARE Score ≤ 4', 'ARE Score > 4'])
    return categories.value_counts()

# Proportion for upregulated genes
up_scores = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score']
up_categories = categorize_scores(up_scores)

# Proportion for downregulated genes
down_scores = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score']
down_categories = categorize_scores(down_scores)

# Set the Seaborn context
sns.set_context("notebook", font_scale=1.25)

# Function to plot pie charts and save them
def plot_and_save_pie_chart(categories, colors, filename, startangle=90, explode=(0, 0, 0.1)):
    fig, ax = plt.subplots()
    def autopct_func(pct, allvals):
        absolute = int(round(pct / 100. * sum(allvals)))
        return "{:.1f}%\n({:d})".format(pct, absolute)

    wedges, texts, autotexts = ax.pie(
        categories.values,
        labels=categories.index,
        colors=colors,
        autopct=lambda pct: autopct_func(pct, categories.values),
        startangle=startangle,
        textprops=dict(color="black"),
        explode=explode,
        wedgeprops={"edgecolor": "black", "linewidth": 1}
    )
    
    for text in autotexts:
        # Move the text higher so it fits better
        text.set_position((text.get_position()[0], text.get_position()[1] - 0.2))
    ax.axis('equal') # Equal aspect ratio ensures that pie is drawn as a circle.
    plt.savefig(filename, format='svg')
    plt.show() # Close the figure to prevent it from displaying immediately and to free memory

# Set a common start angle for consistency
start_angle = 90
# Explode the 'ARE Score = 0' slice larger than the other slices
explode_factor = (0.1, 0, 0)

# Plot and save upregulated pie chart with a gradient from light orange to dark orange
up_colors = ['orange', 'darkorange', 'orangered']
plot_and_save_pie_chart(up_categories, up_colors, 'upregulated_overall_pie.svg', startangle=start_angle, explode=explode_factor)

# Plot and save downregulated pie chart with a gradient from light blue to dark blue
down_colors = ['lightblue', 'blue', 'darkblue']
plot_and_save_pie_chart(down_categories, down_colors, 'downregulated_overall_pie.svg', startangle=start_angle, explode=explode_factor)
Series([], Name: Regulation, dtype: int64)
No description has been provided for this image
No description has been provided for this image

Histogram of ARE score count per ARE score for up and downregulated genes this should show us definitively that we have very few genes with high AREscores in the downreuglated set compared to the upregulated set

In [24]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0] # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Calculate the total number of genes
total_genes = len(combined_data)

# Define bins with a consistent width
binwidth = 1
bins = np.arange(-0.5, max(combined_data['ARE_score']) + binwidth + 0.5, binwidth)

# Plotting the histogram
plt.figure(figsize=(12, 6))

# Normalize the bin counts
upregulated_counts, _ = np.histogram(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'], bins=bins)
downregulated_counts, _ = np.histogram(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'], bins=bins)

# Convert counts to proportions
upregulated_norm = upregulated_counts / total_genes
downregulated_norm = downregulated_counts / total_genes
print(np.sum(upregulated_norm))
print(np.sum(downregulated_norm))
# Upregulated histogram
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'], bins=bins, kde=True, color='orange', label='Upregulated', alpha=0.5, stat='density')
# Downregulated histogram
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'], bins=bins, kde=True, color='blue', label='Downregulated', alpha=0.5, stat='density')
# Labels and Legend
plt.xlabel('ARE Score')
plt.ylabel('Proportion')
# Correctly set the ticks and labels
bin_labels = ['0'] + [f'{int(x)}' for x in np.arange(1, max(combined_data['ARE_score']) + 1)]
ticks = np.arange(0, max(combined_data['ARE_score']) + 1)
plt.xticks(ticks=ticks, labels=bin_labels, rotation=45, ha="center")
plt.title('Distribution of ARE Scores for Upregulated and Downregulated Genes (Normalized)')
plt.legend(title='Regulation')

# Save the figure
plt.savefig("ARE_score_histogram_binned_centered_normalized.svg", format="svg")

# Show the plot
plt.show()
0.5993555316863587
0.40064446831364126
No description has been provided for this image
In [25]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Cap the ARE scores at 20 for any values above 20
combined_data['ARE_score_capped'] = np.where(combined_data['ARE_score'] > 20, 20, combined_data['ARE_score'])

# Define bins as specified, with an additional bin edge for scores above 20
bins = np.concatenate([np.array([0]), np.arange(1, 5.1, 0.1), np.arange(5.1, 10.1, 0.1), np.arange(11, 21, 1), np.array([21])])

# Plotting the histogram
plt.figure(figsize=(12, 6))
# Upregulated histogram
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', label='Upregulated', alpha=0.7, stat='probability')
# Downregulated histogram
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', label='Downregulated', alpha=0.7, stat='probability')

# Labels and Legend
plt.xlabel('ARE Score')
plt.ylabel('Proportion')
# Correctly set the ticks and labels
major_ticks = np.concatenate([np.array([0]), np.arange(1, 6), np.arange(6, 11, 1), np.arange(11, 21, 1)])
major_labels = [str(x) if x != 21 else '20+' for x in major_ticks]
plt.xticks(ticks=major_ticks, labels=major_labels, rotation=45, ha="right")

# Title and Legend
plt.title('Distribution of ARE Scores for Upregulated and Downregulated Genes (Normalized)')
plt.legend(title='Regulation')

# Save the figure
plt.savefig("ARE_score_histogram_custom_bins_normalized.svg", format="svg")
# Show the plot
plt.show()
No description has been provided for this image
In [26]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Cap the ARE scores at 20 for any values above 20
combined_data['ARE_score_capped'] = np.where(combined_data['ARE_score'] > 20, 20, combined_data['ARE_score'])

# Define the bins for each subplot with a step size of 0.1
bins = np.arange(0, 21.1, 0.1)  # Including from 0 to 21 to cover up to 20+

# Create subplots
fig, axes = plt.subplots(2, 2, figsize=(15, 10))

# 0-6
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=axes[0, 0], stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=axes[0, 0], stat='probability', alpha=0.7, label='Downregulated')
axes[0, 0].set_xlim(0, 6)
axes[0, 0].set_title('ARE Scores 0-6')
axes[0, 0].legend(loc='upper right')

# 5-11
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=axes[0, 1], stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=axes[0, 1], stat='probability', alpha=0.7, label='Downregulated')
axes[0, 1].set_xlim(5, 11)
axes[0, 1].set_ylim(0,0.05)
axes[0, 1].set_title('ARE Scores 5-11')
axes[0, 1].legend(loc='upper right')

# 10-16
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=axes[1, 0], stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=axes[1, 0], stat='probability', alpha=0.7, label='Downregulated')
axes[1, 0].set_xlim(10, 16)
axes[1, 0].set_ylim(0,0.05)
axes[1, 0].set_title('ARE Scores 10-16')
axes[1, 0].legend(loc='upper right')

# 15-20+
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=axes[1, 1], stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=axes[1, 1], stat='probability', alpha=0.7, label='Downregulated')
axes[1, 1].set_xlim(15, 20.1)
axes[1, 1].set_ylim(0,0.05)
axes[1, 1].set_title('ARE Scores 15-20+')
axes[1, 1].set_xticks([15, 16, 17, 18, 19, 20])
axes[1, 1].set_xticklabels(['15', '16', '17', '18', '19', '20+'])
axes[1, 1].legend(loc='upper right')

# Set common labels
for ax in axes.flat:
    ax.set_ylabel('Proportion')

# Adjust layout and display
fig.tight_layout()
plt.savefig("ARE_score_histogram_custom_subplots_normalized.svg", format="svg")
plt.show()
No description has been provided for this image
In [27]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0] # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Cap the ARE scores at 20 for any values above 20
combined_data['ARE_score_capped'] = np.where(combined_data['ARE_score'] > 20, 20, combined_data['ARE_score'])

# Define bins for the histogram
bins = np.arange(0, 21.1, 0.1) # Including from 0 to 21 to cover up to 20+

# Plotting the histogram
plt.figure(figsize=(12, 6))
# Upregulated histogram
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', label='Upregulated', alpha=0.7, stat='probability')
# Downregulated histogram
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', label='Downregulated', alpha=0.7, stat='probability')

# Labels and Legend
plt.xlabel('ARE Score')
plt.ylabel('Proportion')
plt.title('Distribution of ARE Scores for Upregulated and Downregulated Genes (Normalized)')

# Define major ticks and labels
major_ticks = np.arange(0, 21, 1)
major_labels = [str(x) if x != 20 else '20+' for x in major_ticks]

# Adjust tick positions and labels
plt.xticks(ticks=major_ticks + 0.05, labels=major_labels, rotation=45, ha="right")

# Title and Legend
plt.legend(title='Regulation')

# Save the figure
plt.savefig("ARE_score_histogram_custom_bins_normalized.svg", format="svg")

# Show the plot
plt.show()
No description has been provided for this image

Redone with no redundant bins

In [28]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Cap the ARE scores at 20 for any values above 20
combined_data['ARE_score_capped'] = np.where(combined_data['ARE_score'] > 20, 20, combined_data['ARE_score'])

# Define the bins for each subplot with a step size of 0.1
bins = np.arange(0, 21.1, 0.1)  # Including from 0 to 21 to cover up to 20+

# Create subplots
fig, axes = plt.subplots(2, 2, figsize=(15, 10))

# 0-5
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=axes[0, 0], stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=axes[0, 0], stat='probability', alpha=0.7, label='Downregulated')
axes[0, 0].set_xlim(0, 5)
axes[0, 0].set_title('ARE Scores 0-5')
axes[0, 0].legend(loc='upper right')

# 5-10
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=axes[0, 1], stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=axes[0, 1], stat='probability', alpha=0.7, label='Downregulated')
axes[0, 1].set_xlim(5, 10)
axes[0, 1].set_title('ARE Scores 5-10')
axes[0, 1].legend(loc='upper right')

# 10-15
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=axes[1, 0], stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=axes[1, 0], stat='probability', alpha=0.7, label='Downregulated')
axes[1, 0].set_xlim(10, 15)
axes[1, 0].set_title('ARE Scores 10-15')
axes[1, 0].legend(loc='upper right')

# 15-20+
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=axes[1, 1], stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=axes[1, 1], stat='probability', alpha=0.7, label='Downregulated')
axes[1, 1].set_xlim(15, 20.1)
axes[1, 1].set_title('ARE Scores 15-20+')
axes[1, 1].set_xticks([15, 16, 17, 18, 19, 20])
axes[1, 1].set_xticklabels(['15', '16', '17', '18', '19', '20+'])
axes[1, 1].legend(loc='upper right')

# Set common labels
for ax in axes.flat:
    ax.set_ylabel('Proportion')

# Adjust layout and display
fig.tight_layout()
plt.savefig("ARE_score_histogram_custom_subplots_normalized.svg", format="svg")
plt.show()
No description has been provided for this image
In [29]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Cap the ARE scores at 20 for any values above 20
combined_data['ARE_score_capped'] = np.where(combined_data['ARE_score'] > 20, 20, combined_data['ARE_score'])

# Define the bins for the histogram with a step size of 0.1
bins = np.arange(0, 21.1, 0.1)

# Create subplots
fig, axes = plt.subplots(2, 2, figsize=(15, 10))

def annotate_bars(ax):
    for p in ax.patches:
        height = p.get_height()
        width = p.get_x() + p.get_width() / 2
        bin_value = round(p.get_x(), 1)  # Annotate the left edge of each bin
        if height > 0:
            ax.annotate(f'{bin_value}', (width, height), ha='center', va='bottom', fontsize=8, rotation=90)

# 0-5
ax = axes[0, 0]
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=ax, stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=ax, stat='probability', alpha=0.7, label='Downregulated')
ax.set_xlim(0, 5)
ax.set_xticks([0, 1, 2, 3, 4, 5])
ax.set_title('ARE Scores 0-5')
ax.legend(loc='upper right')
# annotate_bars(ax)

# 5-10
ax = axes[0, 1]
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=ax, stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=ax, stat='probability', alpha=0.7, label='Downregulated')
ax.set_xlim(5, 10)
ax.set_xticks([5, 6, 7, 8, 9, 10])
ax.set_title('ARE Scores 5-10')
ax.legend(loc='upper right')
# annotate_bars(ax)

# 10-15
ax = axes[1, 0]
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=ax, stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=ax, stat='probability', alpha=0.7, label='Downregulated')
ax.set_xlim(10, 15)
ax.set_xticks([10, 11, 12, 13, 14, 15])
ax.set_title('ARE Scores 10-15')
ax.legend(loc='upper right')
# annotate_bars(ax)

# 15-20+
ax = axes[1, 1]
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=ax, stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=ax, stat='probability', alpha=0.7, label='Downregulated')
ax.set_xlim(15, 20.1)
ax.set_xticks([15, 16, 17, 18, 19, 20])
ax.set_xticklabels(['15', '16', '17', '18', '19', '20+'])
ax.set_title('ARE Scores 15-20+')
ax.legend(loc='upper right')
# annotate_bars(ax)

# Set common labels
for ax in axes.flat:
    ax.set_ylabel('Probability')

# Adjust layout and display
fig.tight_layout()
plt.savefig("ARE_score_histogram_custom_subplots_annotated.svg", format="svg")
plt.show()
No description has been provided for this image
In [30]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Cap the ARE scores at 20 for any values above 20
combined_data['ARE_score_capped'] = np.where(combined_data['ARE_score'] > 20, 20, combined_data['ARE_score'])

# Define the bins for each subplot with a step size of 0.1
bins = np.arange(0, 21.1, 0.1)  # Including from 0 to 21 to cover up to 20+

# First, calculate the overall max proportion for setting consistent y-axis limits
hist_data = sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, stat='probability', alpha=0.7, label='Upregulated')
hist_data = sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, stat='probability', alpha=0.7, label='Downregulated')
max_prop = hist_data.get_ylim()[1]
plt.close()

# Create subplots
fig, axes = plt.subplots(2, 2, figsize=(15, 10))

# 0-5
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=axes[0, 0], stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=axes[0, 0], stat='probability', alpha=0.7, label='Downregulated')
axes[0, 0].set_xlim(0, 5)
axes[0, 0].set_ylim(0, max_prop)
axes[0, 0].set_title('ARE Scores 0-5')
axes[0, 0].legend(loc='upper right')

# 6-10
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=axes[0, 1], stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=axes[0, 1], stat='probability', alpha=0.7, label='Downregulated')
axes[0, 1].set_xlim(6, 10)
axes[0, 1].set_ylim(0, max_prop)
axes[0, 1].set_title('ARE Scores 6-10')
axes[0, 1].legend(loc='upper right')

# 11-15
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=axes[1, 0], stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=axes[1, 0], stat='probability', alpha=0.7, label='Downregulated')
axes[1, 0].set_xlim(11, 15)
axes[1, 0].set_ylim(0, max_prop)
axes[1, 0].set_title('ARE Scores 11-15')
axes[1, 0].legend(loc='upper right')

# 16-20+
sns.histplot(combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score_capped'], bins=bins, kde=False, color='orange', ax=axes[1, 1], stat='probability', alpha=0.7, label='Upregulated')
sns.histplot(combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score_capped'], bins=bins, kde=False, color='blue', ax=axes[1, 1], stat='probability', alpha=0.7, label='Downregulated')
axes[1, 1].set_xlim(16, 20.1)
axes[1, 1].set_ylim(0, max_prop)
axes[1, 1].set_title('ARE Scores 16-20+')
axes[1, 1].set_xticks([16, 17, 18, 19, 20])
axes[1, 1].set_xticklabels(['16', '17', '18', '19', '20+'])
axes[1, 1].legend(loc='upper right')

# Set common labels
for ax in axes.flat:
    ax.set_ylabel('Proportion')

# Adjust layout and display
fig.tight_layout()
plt.savefig("ARE_score_histogram_custom_subplots_normalized.svg", format="svg")
plt.show()
No description has been provided for this image
In [31]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0] # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score'])
combined_data = combined_data[combined_data['ARE_score'] >= 0]

# Check for any ARE scores above 20
scores_above_20 = combined_data[combined_data['ARE_score'] > 20]

# Print the number of scores above 20
print(f"Number of ARE scores above 20: {len(scores_above_20)}")
Number of ARE scores above 20: 49

Here is ARE-score now plotted against log2fc as a simple volcano plot:

In [32]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score', 'log2FoldChange'])

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Plotting the volcano plot
plt.figure(figsize=(12, 8))

# Volcano plot
sns.scatterplot(data=combined_data, x='log2FoldChange', y='ARE_score', hue='Regulation', alpha=0.7, palette={'Upregulated': 'orange', 'Downregulated': 'blue'})

# Labels and Title
plt.xlabel('log2FoldChange')
plt.ylabel('ARE Score')
plt.title('Volcano Plot of ARE Scores vs log2FoldChange')

# Adding horizontal and vertical lines to separate upregulated and downregulated genes if required
plt.axvline(x=0, linestyle='--', color='black', linewidth=1)

# Show Legend
plt.legend(title='Regulation')

# Save the figure
plt.savefig("ARE_score_vs_log2FoldChange_volcano_plot.svg", format="svg")

# Show the plot
plt.show()
No description has been provided for this image

Here I add some marginal histograms for both variables as another way to associate these variables and their trends between them

In [33]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os
from scipy.stats import pearsonr

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score', 'log2FoldChange'])

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Setting up the figure with marginal histograms using JointGrid
g = sns.JointGrid(data=combined_data, x='log2FoldChange', y='ARE_score', height=10)

# Add scatterplot
sns.scatterplot(data=combined_data, x='log2FoldChange', y='ARE_score', hue='Regulation', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_joint, alpha=0.6)

# Plot trend lines
sns.regplot(data=combined_data[combined_data['Regulation'] == 'Upregulated'], x='log2FoldChange', y='ARE_score', scatter=False, ax=g.ax_joint, color='orange', label='Upregulated Trend')
sns.regplot(data=combined_data[combined_data['Regulation'] == 'Downregulated'], x='log2FoldChange', y='ARE_score', scatter=False, ax=g.ax_joint, color='blue', label='Downregulated Trend')

# Calculate and annotate Pearson correlation coefficient
pearson_corr_up, _ = pearsonr(combined_data[combined_data['Regulation'] == 'Upregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'])
pearson_corr_down, _ = pearsonr(combined_data[combined_data['Regulation'] == 'Downregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'])

# Add annotations for Pearson correlation
g.ax_joint.text(0.05, 0.95, f'Upreg. Pearson r: {pearson_corr_up:.2f}', transform=g.ax_joint.transAxes, color='orange', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.05, 0.90, f'Downreg. Pearson r: {pearson_corr_down:.2f}', transform=g.ax_joint.transAxes, color='blue', fontsize=12, verticalalignment='top')

# Add titles and labels
g.set_axis_labels("log2FoldChange", "ARE Score")
g.fig.suptitle('Joint Plot of ARE Scores vs log2FoldChange with Marginal Histograms and Trend Lines', y=1.03)

# Customize marginal distributions for better visibility, separate by 'Regulation'
sns.histplot(data=combined_data, x='log2FoldChange', hue='Regulation', multiple='stack', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_x, bins=30, alpha=0.6)
sns.histplot(data=combined_data, y='ARE_score', hue='Regulation', multiple='stack', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_y, bins=30, alpha=0.6)

# Add legend for trend lines
g.ax_joint.legend(loc='upper left')

# Save the figure
g.savefig("ARE_score_vs_log2FoldChange_joint_plot_annotated_improved.svg", format="svg")

# Show the plot
plt.show()
No description has been provided for this image

Same thing with mean and median annotations added and legends so that they do not overlap one another

In [34]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os
from scipy.stats import pearsonr

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0] # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score', 'log2FoldChange'])

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Setting up the figure with marginal histograms using JointGrid
g = sns.JointGrid(data=combined_data, x='log2FoldChange', y='ARE_score', height=10)

# Add scatterplot without legend
sns.scatterplot(data=combined_data, x='log2FoldChange', y='ARE_score', hue='Regulation', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_joint, alpha=0.6, legend=False)

# Plot trend lines
up_trend = sns.regplot(data=combined_data[combined_data['Regulation'] == 'Upregulated'], x='log2FoldChange', y='ARE_score', scatter=False, ax=g.ax_joint, color='orange', label='Upregulated Trend')
down_trend = sns.regplot(data=combined_data[combined_data['Regulation'] == 'Downregulated'], x='log2FoldChange', y='ARE_score', scatter=False, ax=g.ax_joint, color='blue', label='Downregulated Trend')

# Calculate and annotate Pearson correlation coefficient
pearson_corr_up, _ = pearsonr(combined_data[combined_data['Regulation'] == 'Upregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'])
pearson_corr_down, _ = pearsonr(combined_data[combined_data['Regulation'] == 'Downregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'])

# Add annotations for Pearson correlation
g.ax_joint.text(0.05, 0.95, f'Upreg. Pearson r: {pearson_corr_up:.2f}', transform=g.ax_joint.transAxes, color='orange', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.05, 0.90, f'Downreg. Pearson r: {pearson_corr_down:.2f}', transform=g.ax_joint.transAxes, color='blue', fontsize=12, verticalalignment='top')

# Add horizontal lines to indicate threshold in ARE scores
g.ax_joint.axhline(y=0, color='grey', linestyle='--', linewidth=1)

# Add titles and labels
g.set_axis_labels("log2FoldChange", "ARE Score")
g.fig.suptitle('Joint Plot of ARE Scores vs log2FoldChange with Marginal Histograms and Trend Lines', y=1.03)

# Customize marginal distributions for better visibility, separate by 'Regulation'
sns.histplot(data=combined_data, x='log2FoldChange', hue='Regulation', multiple='stack', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_x, bins=30, alpha=0.6)
sns.histplot(data=combined_data, y='ARE_score', hue='Regulation', multiple='stack', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_y, bins=30, alpha=0.6)

# Add density plots on top of the histograms to show the distribution more clearly
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Upregulated'], x='log2FoldChange', ax=g.ax_marg_x, color='orange', fill=True, alpha=0.2)
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Downregulated'], x='log2FoldChange', ax=g.ax_marg_x, color='blue', fill=True, alpha=0.2)
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Upregulated'], y='ARE_score', ax=g.ax_marg_y, color='orange', fill=True, alpha=0.2)
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Downregulated'], y='ARE_score', ax=g.ax_marg_y, color='blue', fill=True, alpha=0.2)

# Add summary statistics as text annotations
mean_up = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'].mean()
mean_down = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'].mean()
median_up = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'].median()
median_down = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'].median()
g.ax_joint.text(0.65, 0.85, f'Upreg. ARE Mean: {mean_up:.2f}\nUpreg. ARE Median: {median_up:.2f}', transform=g.ax_joint.transAxes, color='orange', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.65, 0.75, f'Downreg. ARE Mean: {mean_down:.2f}\nDownreg. ARE Median: {median_down:.2f}', transform=g.ax_joint.transAxes, color='blue', fontsize=12, verticalalignment='top')

# Add a single custom legend
handles, labels = g.ax_joint.get_legend_handles_labels()
g.ax_joint.legend(handles=handles, labels=["Upregulated", "Downregulated", "Upregulated Trend", "Downregulated Trend"], loc="upper right", frameon=True)

# Save the figure
g.savefig("ARE_score_vs_log2FoldChange_joint_plot_annotated_improved_further.svg", format="svg")

# Show the plot
plt.show()
/tmp/ipykernel_2823251/2894176590.py:80: UserWarning: Mismatched number of handles and labels: len(handles) = 2 len(labels) = 4
  g.ax_joint.legend(handles=handles, labels=["Upregulated", "Downregulated", "Upregulated Trend", "Downregulated Trend"], loc="upper right", frameon=True)
No description has been provided for this image

Here is the same plot with slope annotations added

In [35]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os
from scipy.stats import pearsonr

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score', 'log2FoldChange'])

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Setting up the figure with marginal histograms using JointGrid
g = sns.JointGrid(data=combined_data, x='log2FoldChange', y='ARE_score', height=10)

# Add scatterplot without legend
sns.scatterplot(data=combined_data, x='log2FoldChange', y='ARE_score', hue='Regulation', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_joint, alpha=0.6, legend=False)

# Plot trend lines
up_trend = sns.regplot(data=combined_data[combined_data['Regulation'] == 'Upregulated'], x='log2FoldChange', y='ARE_score', scatter=False, ax=g.ax_joint, color='orange', label='Upregulated Trend')
down_trend = sns.regplot(data=combined_data[combined_data['Regulation'] == 'Downregulated'], x='log2FoldChange', y='ARE_score', scatter=False, ax=g.ax_joint, color='blue', label='Downregulated Trend')

# Calculate and annotate Pearson correlation coefficient
pearson_corr_up, _ = pearsonr(combined_data[combined_data['Regulation'] == 'Upregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'])
pearson_corr_down, _ = pearsonr(combined_data[combined_data['Regulation'] == 'Downregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'])

# Calculate slopes for trend lines
slope_up = np.polyfit(combined_data[combined_data['Regulation'] == 'Upregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'], 1)[0]
slope_down = np.polyfit(combined_data[combined_data['Regulation'] == 'Downregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'], 1)[0]

# Add annotations for Pearson correlation and slopes
g.ax_joint.text(0.05, 0.95, f'Upreg. Pearson r: {pearson_corr_up:.2f}', transform=g.ax_joint.transAxes, color='orange', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.05, 0.90, f'Downreg. Pearson r: {pearson_corr_down:.2f}', transform=g.ax_joint.transAxes, color='blue', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.05, 0.85, f'Upreg. Slope: {slope_up:.3f}', transform=g.ax_joint.transAxes, color='orange', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.05, 0.80, f'Downreg. Slope: {slope_down:.3f}', transform=g.ax_joint.transAxes, color='blue', fontsize=12, verticalalignment='top')

# Add horizontal lines to indicate threshold in ARE scores
g.ax_joint.axhline(y=0, color='grey', linestyle='--', linewidth=1)

# Add titles and labels
g.set_axis_labels("log2FoldChange", "ARE Score")
g.fig.suptitle('Joint Plot of ARE Scores vs log2FoldChange with Marginal Histograms and Trend Lines', y=1.03)

# Customize marginal distributions for better visibility, separate by 'Regulation'
sns.histplot(data=combined_data, x='log2FoldChange', hue='Regulation', multiple='stack', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_x, bins=30, alpha=0.6)
sns.histplot(data=combined_data, y='ARE_score', hue='Regulation', multiple='stack', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_y, bins=30, alpha=0.6)

# Add density plots on top of the histograms to show the distribution more clearly
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Upregulated'], x='log2FoldChange', ax=g.ax_marg_x, color='orange', fill=True, alpha=0.2)
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Downregulated'], x='log2FoldChange', ax=g.ax_marg_x, color='blue', fill=True, alpha=0.2)
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Upregulated'], y='ARE_score', ax=g.ax_marg_y, color='orange', fill=True, alpha=0.2)
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Downregulated'], y='ARE_score', ax=g.ax_marg_y, color='blue', fill=True, alpha=0.2)

# Add summary statistics as text annotations
mean_up = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'].mean()
mean_down = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'].mean()
median_up = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'].median()
median_down = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'].median()
g.ax_joint.text(0.65, 0.85, f'Upreg. ARE Mean: {mean_up:.2f}\nUpreg. ARE Median: {median_up:.2f}', transform=g.ax_joint.transAxes, color='orange', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.65, 0.75, f'Downreg. ARE Mean: {mean_down:.2f}\nDownreg. ARE Median: {median_down:.2f}', transform=g.ax_joint.transAxes, color='blue', fontsize=12, verticalalignment='top')

# Add a single custom legend
handles, labels = g.ax_joint.get_legend_handles_labels()
g.ax_joint.legend(handles=handles, labels=["Upregulated", "Downregulated", "Upregulated Trend", "Downregulated Trend"], loc="upper right", frameon=True)

# Save the figure
g.savefig("ARE_score_vs_log2FoldChange_joint_plot_annotated_slopes.svg", format="svg")

# Show the plot
plt.show()
/tmp/ipykernel_2823251/2272620246.py:86: UserWarning: Mismatched number of handles and labels: len(handles) = 2 len(labels) = 4
  g.ax_joint.legend(handles=handles, labels=["Upregulated", "Downregulated", "Upregulated Trend", "Downregulated Trend"], loc="upper right", frameon=True)
No description has been provided for this image

Same plot but this time I use density hexgram

In [36]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os
from scipy.stats import pearsonr

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score', 'log2FoldChange'])

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Setting up the figure with marginal histograms using JointGrid
g = sns.JointGrid(data=combined_data, x='log2FoldChange', y='ARE_score', height=10)

# Add hexbin plot to show density
hb = g.ax_joint.hexbin(combined_data['log2FoldChange'], combined_data['ARE_score'], gridsize=50, cmap='Blues', mincnt=1)

# Add color bar for the hexbin plot
cb = g.fig.colorbar(hb, ax=g.ax_joint, orientation='vertical')
cb.set_label('Counts')

# Plot trend lines
up_trend = sns.regplot(data=combined_data[combined_data['Regulation'] == 'Upregulated'], x='log2FoldChange', y='ARE_score', scatter=False, ax=g.ax_joint, color='orange', label='Upregulated Trend')
down_trend = sns.regplot(data=combined_data[combined_data['Regulation'] == 'Downregulated'], x='log2FoldChange', y='ARE_score', scatter=False, ax=g.ax_joint, color='blue', label='Downregulated Trend')

# Calculate and annotate Pearson correlation coefficient
pearson_corr_up, _ = pearsonr(combined_data[combined_data['Regulation'] == 'Upregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'])
pearson_corr_down, _ = pearsonr(combined_data[combined_data['Regulation'] == 'Downregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'])

# Calculate slopes for trend lines
slope_up = np.polyfit(combined_data[combined_data['Regulation'] == 'Upregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'], 1)[0]
slope_down = np.polyfit(combined_data[combined_data['Regulation'] == 'Downregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'], 1)[0]

# Add annotations for Pearson correlation and slopes
g.ax_joint.text(0.05, 0.95, f'Upreg. Pearson r: {pearson_corr_up:.2f}', transform=g.ax_joint.transAxes, color='orange', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.05, 0.90, f'Downreg. Pearson r: {pearson_corr_down:.2f}', transform=g.ax_joint.transAxes, color='blue', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.05, 0.85, f'Upreg. Slope: {slope_up:.3f}', transform=g.ax_joint.transAxes, color='orange', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.05, 0.80, f'Downreg. Slope: {slope_down:.3f}', transform=g.ax_joint.transAxes, color='blue', fontsize=12, verticalalignment='top')

# Add horizontal lines to indicate threshold in ARE scores
g.ax_joint.axhline(y=0, color='grey', linestyle='--', linewidth=1)

# Add titles and labels
g.set_axis_labels("log2FoldChange", "ARE Score")
g.fig.suptitle('Joint Plot of ARE Scores vs log2FoldChange with Hexbin Density and Trend Lines', y=1.03)

# Customize marginal distributions for better visibility, separate by 'Regulation'
sns.histplot(data=combined_data, x='log2FoldChange', hue='Regulation', multiple='stack', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_x, bins=30, alpha=0.6)
sns.histplot(data=combined_data, y='ARE_score', hue='Regulation', multiple='stack', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_y, bins=30, alpha=0.6)

# Add summary statistics as text annotations
mean_up = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'].mean()
mean_down = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'].mean()
median_up = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'].median()
median_down = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'].median()
g.ax_joint.text(0.65, 0.85, f'Upreg. ARE Mean: {mean_up:.2f}\nUpreg. ARE Median: {median_up:.2f}', transform=g.ax_joint.transAxes, color='orange', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.65, 0.75, f'Downreg. ARE Mean: {mean_down:.2f}\nDownreg. ARE Median: {median_down:.2f}', transform=g.ax_joint.transAxes, color='blue', fontsize=12, verticalalignment='top')

# Add a single custom legend
handles, labels = g.ax_joint.get_legend_handles_labels()
g.ax_joint.legend(handles=handles, labels=["Upregulated", "Downregulated", "Upregulated Trend", "Downregulated Trend"], loc="upper right", frameon=True)

# Save the figure
g.savefig("ARE_score_vs_log2FoldChange_joint_plot_annotated_density.svg", format="svg")

# Show the plot
plt.show()
/tmp/ipykernel_2823251/1615529175.py:84: UserWarning: Mismatched number of handles and labels: len(handles) = 2 len(labels) = 4
  g.ax_joint.legend(handles=handles, labels=["Upregulated", "Downregulated", "Upregulated Trend", "Downregulated Trend"], loc="upper right", frameon=True)
No description has been provided for this image

Now here is an iteration of this same plot just this time all genes must have AREscore>0 and a log2FC absolute value of 2 or more these can be modified to watch the distribution change

In [37]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os
from scipy.stats import pearsonr

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Loading the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combining the data and excluding rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score', 'log2FoldChange'])

# Filtering to include only genes with ARE scores greater than 0 and absolute value of log2FoldChange greater than 2
combined_data = combined_data[(combined_data['ARE_score'] > 0) & (combined_data['log2FoldChange'].abs() > 4)]

# Dropping duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Setting up the figure with marginal histograms using JointGrid
g = sns.JointGrid(data=combined_data, x='log2FoldChange', y='ARE_score', height=10)

# Add scatterplot without legend
sns.scatterplot(data=combined_data, x='log2FoldChange', y='ARE_score', hue='Regulation', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_joint, alpha=0.6, legend=False)

# Plot trend lines
up_trend = sns.regplot(data=combined_data[combined_data['Regulation'] == 'Upregulated'], x='log2FoldChange', y='ARE_score', scatter=False, ax=g.ax_joint, color='orange', label='Upregulated Trend')
down_trend = sns.regplot(data=combined_data[combined_data['Regulation'] == 'Downregulated'], x='log2FoldChange', y='ARE_score', scatter=False, ax=g.ax_joint, color='blue', label='Downregulated Trend')

# Calculate and annotate Pearson correlation coefficient
pearson_corr_up, _ = pearsonr(combined_data[combined_data['Regulation'] == 'Upregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'])
pearson_corr_down, _ = pearsonr(combined_data[combined_data['Regulation'] == 'Downregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'])

# Calculate slopes for trend lines
slope_up = np.polyfit(combined_data[combined_data['Regulation'] == 'Upregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'], 1)[0]
slope_down = np.polyfit(combined_data[combined_data['Regulation'] == 'Downregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'], 1)[0]

# Add annotations for Pearson correlation and slopes
g.ax_joint.text(0.05, 0.95, f'Upreg. Pearson r: {pearson_corr_up:.2f}', transform=g.ax_joint.transAxes, color='orange', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.05, 0.90, f'Downreg. Pearson r: {pearson_corr_down:.2f}', transform=g.ax_joint.transAxes, color='blue', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.05, 0.85, f'Upreg. Slope: {slope_up:.3f}', transform=g.ax_joint.transAxes, color='orange', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.05, 0.80, f'Downreg. Slope: {slope_down:.3f}', transform=g.ax_joint.transAxes, color='blue', fontsize=12, verticalalignment='top')

# Add horizontal lines to indicate threshold in ARE scores
g.ax_joint.axhline(y=0, color='grey', linestyle='--', linewidth=1)

# Add titles and labels
g.set_axis_labels("log2FoldChange", "ARE Score")
g.fig.suptitle('Joint Plot of ARE Scores vs log2FoldChange with Marginal Histograms and Trend Lines', y=1.03)

# Customize marginal distributions for better visibility, separate by 'Regulation'
sns.histplot(data=combined_data, x='log2FoldChange', hue='Regulation', multiple='stack', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_x, bins=30, alpha=0.6)
sns.histplot(data=combined_data, y='ARE_score', hue='Regulation', multiple='stack', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_y, bins=30, alpha=0.6)

# Add density plots on top of the histograms to show the distribution more clearly
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Upregulated'], x='log2FoldChange', ax=g.ax_marg_x, color='orange', fill=True, alpha=0.2)
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Downregulated'], x='log2FoldChange', ax=g.ax_marg_x, color='blue', fill=True, alpha=0.2)
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Upregulated'], y='ARE_score', ax=g.ax_marg_y, color='orange', fill=True, alpha=0.2)
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Downregulated'], y='ARE_score', ax=g.ax_marg_y, color='blue', fill=True, alpha=0.2)

# Add summary statistics as text annotations
mean_up = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'].mean()
mean_down = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'].mean()
median_up = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'].median()
median_down = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'].median()
g.ax_joint.text(0.65, 0.85, f'Upreg. ARE Mean: {mean_up:.2f}\nUpreg. ARE Median: {median_up:.2f}', transform=g.ax_joint.transAxes, color='orange', fontsize=12, verticalalignment='top')
g.ax_joint.text(0.65, 0.75, f'Downreg. ARE Mean: {mean_down:.2f}\nDownreg. ARE Median: {median_down:.2f}', transform=g.ax_joint.transAxes, color='blue', fontsize=12, verticalalignment='top')

# Add a single custom legend
handles, labels = g.ax_joint.get_legend_handles_labels()
g.ax_joint.legend(handles=handles, labels=["Upregulated", "Downregulated", "Upregulated Trend", "Downregulated Trend"], loc="upper right", frameon=True)

# Save the figure
g.savefig("ARE_score_vs_log2FoldChange_joint_plot_annotated_filtered_log2fc.svg", format="svg")

# Show the plot
plt.show()
/tmp/ipykernel_2823251/236085898.py:89: UserWarning: Mismatched number of handles and labels: len(handles) = 2 len(labels) = 4
  g.ax_joint.legend(handles=handles, labels=["Upregulated", "Downregulated", "Upregulated Trend", "Downregulated Trend"], loc="upper right", frameon=True)
No description has been provided for this image

Now I do this same plot as a hexbin plot to see if it does this better

In [38]:
plt.figure(figsize=(12, 8))
plt.hexbin(combined_data['log2FoldChange'], combined_data['ARE_score'], gridsize=50, cmap='viridis', extent=(-10, 10, 0, 60))
cb = plt.colorbar(label='Count')
plt.title('Hexbin plot of ARE Scores vs log2FoldChange')
plt.xlabel('log2FoldChange')
plt.ylabel('ARE Score')
plt.savefig('hexbin_plot_ARE_score_vs_log2FoldChange.svg', format='svg')
plt.show()
No description has been provided for this image

Here is an optimized hexbin plot

In [39]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os
from scipy.stats import pearsonr

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Load the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and exclude rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score', 'log2FoldChange'])

# Drop duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Create hexbin plot
plt.figure(figsize=(14, 10))
hb = plt.hexbin(combined_data['log2FoldChange'], combined_data['ARE_score'], gridsize=70, cmap='viridis', extent=(-10, 10, 0, 50), mincnt=1)
cb = plt.colorbar(label='Count')
cb.set_label('Count', fontsize=14)

# Add mean ARE scores for upregulated and downregulated
mean_upreg_are = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'].mean()
mean_downreg_are = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'].mean()
plt.axvline(0, color='black', linestyle='--', linewidth=2)
plt.text(4, 45, f'Upreg. Mean ARE: {mean_upreg_are:.2f}', color='orange', fontsize=14, bbox=dict(facecolor='white', alpha=0.7))
plt.text(-9, 45, f'Downreg. Mean ARE: {mean_downreg_are:.2f}', color='blue', fontsize=14, bbox=dict(facecolor='white', alpha=0.7))

# Annotate dense regions
densest_bin = hb.get_offsets()[np.argmax(hb.get_array())]
plt.annotate('Most Dense Region', xy=densest_bin, xytext=(densest_bin[0], densest_bin[1]+5),
             arrowprops=dict(facecolor='white', shrink=0.05), color='white', fontsize=12, ha='center', bbox=dict(facecolor='black', alpha=0.5))

# Add titles and labels
plt.title('Enhanced Hexbin Plot of ARE Scores vs log2FoldChange', fontsize=18, pad=20)
plt.xlabel('log2FoldChange', fontsize=14)
plt.ylabel('ARE Score', fontsize=14)

# Add grid lines and set axis limits
plt.grid(True, which='both', linestyle='--', linewidth=0.7, alpha=0.7)
plt.xlim(-10, 10)
plt.ylim(0, 50)

# Save the figure
plt.savefig('enhanced_hexbin_plot_ARE_score_vs_log2FoldChange_fullrange.svg', format='svg')

# Show the plot
plt.show()
No description has been provided for this image

Here is the scatter plot but instead of lines of best fit here I use density annotations

In [40]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os
from scipy.stats import pearsonr

# Function to load data from CSV files
def load_data(directory, regulation_type):
    all_data = []
    for filename in os.listdir(directory):
        if filename.endswith('_updated.csv'):
            file_path = os.path.join(directory, filename)
            data = pd.read_csv(file_path)
            data['Regulation'] = regulation_type
            data['Tissue'] = filename.split('_')[0]  # Assuming the filename contains the tissue name at the start
            all_data.append(data)
    return pd.concat(all_data, ignore_index=True)

# Define the paths to your data directories, adjust these paths to your actual dataset paths
upregulated_path = './upregulated_genes_updated'
downregulated_path = './downregulated_genes_updated'

# Load the upregulated and downregulated data
upregulated_data = load_data(upregulated_path, 'Upregulated')
downregulated_data = load_data(downregulated_path, 'Downregulated')

# Combine the data and exclude rows with Score = 0 or missing
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score', 'log2FoldChange'])

# Drop duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Set up the figure with marginal histograms using JointGrid
g = sns.JointGrid(data=combined_data, x='log2FoldChange', y='ARE_score', height=10)

# Add scatterplot with reduced alpha to make points more distinct
sns.scatterplot(data=combined_data, x='log2FoldChange', y='ARE_score', hue='Regulation', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_joint, alpha=0.3)

# Add density plot overlay
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Upregulated'], x='log2FoldChange', y='ARE_score', ax=g.ax_joint, color='orange', alpha=0.5, linewidths=1.5)
sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Downregulated'], x='log2FoldChange', y='ARE_score', ax=g.ax_joint, color='blue', alpha=0.5, linewidths=1.5)

# Calculate and annotate Pearson correlation coefficient
pearson_corr_up, _ = pearsonr(combined_data[combined_data['Regulation'] == 'Upregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'])
pearson_corr_down, _ = pearsonr(combined_data[combined_data['Regulation'] == 'Downregulated']['log2FoldChange'], combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'])

# Add annotations for Pearson correlation
# Adjust positions to prevent overlap
g.ax_joint.text(-9.5, 50, f'Upreg. Pearson r: {pearson_corr_up:.2f}', color='orange', fontsize=12)
g.ax_joint.text(-9.5, 45, f'Downreg. Pearson r: {pearson_corr_down:.2f}', color='blue', fontsize=12)

# Calculate mean ARE scores and annotate
mean_upreg_are = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'].mean()
mean_downreg_are = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'].mean()

# Adjust positions to prevent overlap
g.ax_joint.text(4.5, 50, f'Upreg. Mean ARE: {mean_upreg_are:.2f}', color='orange', fontsize=12)
g.ax_joint.text(4.5, 45, f'Downreg. Mean ARE: {mean_downreg_are:.2f}', color='blue', fontsize=12)

# Add Upregulated and Downregulated labels
g.ax_joint.text(3, 40, 'Upregulated', fontsize=14, color='orange', weight='bold')
g.ax_joint.text(-9, 40, 'Downregulated', fontsize=14, color='blue', weight='bold')

# Add titles and labels
g.set_axis_labels("log2FoldChange", "ARE Score")
g.fig.suptitle('Joint Plot of ARE Scores vs log2FoldChange with Marginal Histograms, Trend Lines, and Density', y=1.03)

# Customize marginal distributions for better visibility, separate by 'Regulation'
sns.histplot(data=combined_data, x='log2FoldChange', hue='Regulation', multiple='stack', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_x, bins=30, alpha=0.6, legend=False)
sns.histplot(data=combined_data, y='ARE_score', hue='Regulation', multiple='stack', palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_y, bins=30, alpha=0.6, legend=False)

# Move joint plot and legend only for main plot
handles, labels = g.ax_joint.get_legend_handles_labels()
g.ax_joint.legend(handles=handles, labels=labels, loc='upper left', bbox_to_anchor=(1, 1.2), title='Regulation')

# Save the figure
g.savefig("ARE_score_vs_log2FoldChange_joint_plot_with_density_improved.svg", format="svg")

# Show the plot
plt.show()
No description has been provided for this image

Density annotations with an overall line of best fit which kind of shows that there is an ARE score increase across up from downregulated

In [41]:
import matplotlib.pyplot as plt
import seaborn as sns
import pandas as pd
import numpy as np
import os
from scipy.stats import pearsonr, ttest_ind

# Load data - Your existing code here

# Calculate mean ARE scores and T-test p-value
mean_upreg_are = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'].mean()
mean_downreg_are = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'].mean()

# T-test to compare ARE scores
t_stat, p_value = ttest_ind(
    combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'],
    combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score']
)

# Code for the plot
g = sns.JointGrid(data=combined_data, x='log2FoldChange', y='ARE_score', height=10)

sns.scatterplot(data=combined_data, x='log2FoldChange', y='ARE_score', hue='Regulation', 
                palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_joint, alpha=0.3)

sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Upregulated'], x='log2FoldChange', y='ARE_score', 
            ax=g.ax_joint, color='orange', alpha=0.5, linewidths=1.5)

sns.kdeplot(data=combined_data[combined_data['Regulation'] == 'Downregulated'], x='log2FoldChange', y='ARE_score', 
            ax=g.ax_joint, color='blue', alpha=0.5, linewidths=1.5)

sns.histplot(data=combined_data, x='log2FoldChange', hue='Regulation', multiple='stack', 
             palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_x, bins=30, alpha=0.6, legend=False)

sns.histplot(data=combined_data, y='ARE_score', hue='Regulation', multiple='stack', 
             palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, ax=g.ax_marg_y, bins=30, alpha=0.6, legend=False)

g.ax_joint.text(4.5, 50, f'Upreg. Mean ARE: {mean_upreg_are:.2f}', color='orange', fontsize=12)
g.ax_joint.text(4.5, 45, f'Downreg. Mean ARE: {mean_downreg_are:.2f}', color='blue', fontsize=12)

# Add t-test p-value
g.ax_joint.text(4.5, 40, f'T-test p-value: {p_value:.2e}', color='green', fontsize=12)

sns.regplot(data=combined_data, x='log2FoldChange', y='ARE_score', scatter=False, ax=g.ax_joint, line_kws={"color":"red"})

g.set_axis_labels("log2FoldChange", "ARE Score")
g.fig.suptitle('Joint Plot of ARE Scores vs log2FoldChange with Marginal Histograms, Regression Line, and Density', y=1.03)

handles, labels = g.ax_joint.get_legend_handles_labels()
g.ax_joint.legend(handles=handles, labels=labels, loc='upper left', bbox_to_anchor=(1, 1.2), title='Regulation')

g.savefig("ARE_score_vs_log2FoldChange_joint_plot_with_density_regression_ttest.svg", format="svg")
plt.show()

# Boxplot comparison
plt.figure(figsize=(10, 6))
sns.boxplot(data=combined_data, x='Regulation', y='ARE_score', palette={'Upregulated': 'orange', 'Downregulated': 'blue'})
plt.title('Boxplot of ARE Scores for Upregulated and Downregulated Genes')
plt.savefig("ARE_score_boxplot.svg", format="svg")
plt.show()
No description has been provided for this image
/tmp/ipykernel_2823251/3130667382.py:57: FutureWarning: 

Passing `palette` without assigning `hue` is deprecated and will be removed in v0.14.0. Assign the `x` variable to `hue` and set `legend=False` for the same effect.

  sns.boxplot(data=combined_data, x='Regulation', y='ARE_score', palette={'Upregulated': 'orange', 'Downregulated': 'blue'})
No description has been provided for this image

This plot was made to highlight the tail ends of the distributions

In [42]:
g = sns.FacetGrid(combined_data, col="Regulation", palette={'Upregulated': 'orange', 'Downregulated': 'blue'}, height=6, aspect=1.2)
g.map(sns.scatterplot, "log2FoldChange", "ARE_score", alpha=0.3)
g.map(sns.lineplot, "log2FoldChange", "ARE_score", ci=None, estimator=np.median)

g.set_axis_labels("log2FoldChange", "ARE Score")
g.add_legend()
plt.subplots_adjust(top=0.8)
g.fig.suptitle('Scatter Plots of ARE Scores vs log2FoldChange for Upregulated and Downregulated Genes')
g.savefig("ARE_score_facet_scatter_plot.svg", format="svg")
plt.show()
/home/gsgeorge/.conda/envs/mus_musculus/lib/python3.12/site-packages/seaborn/axisgrid.py:854: FutureWarning: 

The `ci` parameter is deprecated. Use `errorbar=None` for the same effect.

  func(*plot_args, **plot_kwargs)
/home/gsgeorge/.conda/envs/mus_musculus/lib/python3.12/site-packages/seaborn/axisgrid.py:854: FutureWarning: 

The `ci` parameter is deprecated. Use `errorbar=None` for the same effect.

  func(*plot_args, **plot_kwargs)
No description has been provided for this image

Better annotated version of the plot above:

In [43]:
import seaborn as sns
import matplotlib.pyplot as plt
import pandas as pd
import numpy as np

# Sample data loading function here
# all_data = ... (This represents your loaded data)

# Combine the data and exclude rows with ARE_score = 0 or missing values
combined_data = pd.concat([upregulated_data, downregulated_data], ignore_index=True)
combined_data = combined_data.dropna(subset=['ARE_score', 'log2FoldChange'])

# Drop duplicate IDs within the same tissue for up and downregulated genes
combined_data = combined_data.drop_duplicates(subset=['ID', 'Tissue', 'Regulation'])

# Calculate mean ARE scores for annotation
mean_upreg_are = combined_data[combined_data['Regulation'] == 'Upregulated']['ARE_score'].mean()
mean_downreg_are = combined_data[combined_data['Regulation'] == 'Downregulated']['ARE_score'].mean()

# Setting up the FacetGrid
palette = {'Upregulated': 'orange', 'Downregulated': 'blue'}
g = sns.FacetGrid(combined_data, col="Regulation", palette=palette, height=6, aspect=1.2)

# Adding scatter plot
g.map(sns.scatterplot, "log2FoldChange", "ARE_score", alpha=0.3, edgecolor=None)

# Adding smooth trend lines
g.map(sns.regplot, "log2FoldChange", "ARE_score", scatter=False, ci=None, lowess=True, color='red')

# Adding mean ARE score annotations
def annotate(data, **kws):
    reg = data['Regulation'].iloc[0]
    ax = plt.gca()
    if reg == 'Upregulated':
        mean_are = mean_upreg_are
        color = 'orange'
    else:
        mean_are = mean_downreg_are
        color = 'blue'
    ax.text(0.05, 0.95, f'Mean ARE: {mean_are:.2f}', transform=ax.transAxes, color=color, fontsize=12, verticalalignment='top')

g.map_dataframe(annotate)

# Adjusting the axes labels and title
g.set_axis_labels("log2FoldChange", "ARE Score")
g.fig.suptitle('Scatter Plots of ARE Scores vs log2FoldChange for Upregulated and Downregulated Genes', y=1.02)

# Adjusting layout for better appearance
plt.subplots_adjust(top=0.85, wspace=0.3)

# Save the figure
g.savefig("ARE_score_facet_scatter_plot_annotated.svg", format="svg")

# Show the plot
plt.show()
No description has been provided for this image
In [44]:
import os
import pandas as pd

# Function to search for a list of gene names within CSV files in directories
def find_genes_in_csv(directories, gene_names):
    gene_files = {gene: [] for gene in gene_names}
    
    for directory in directories:
        for root, _, files in os.walk(directory):
            csv_files = [f for f in files if f.endswith('.csv')]
            for file in csv_files:
                file_path = os.path.join(root, file)
                try:
                    # Read the CSV file into a DataFrame
                    df = pd.read_csv(file_path)
                    
                    # Check if Gene.name column exists
                    if 'Gene.name' in df.columns:
                        for gene in gene_names:
                            if gene in df['Gene.name'].values:
                                gene_files[gene].append(file_path)
                except Exception as e:
                    print(f"Error reading {file}: {e}")

    return gene_files

# List of directories to search
directories = [
    '/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/upregulated_genes_updated',
    '/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/downregulated_genes_updated',
    '/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated'
]

# List of genes to search for
gene_names_of_interest = [
    'Notch1', 
    'Cebpb',  # CCAAT/enhancer binding protein-beta
    'Junb', 
    'Il1b', 
    'Klf2', 
    'Zfp36', 
    'Zfp36l1', 
    'Itgb2'
]

# Find CSV files containing the gene names
gene_files = find_genes_in_csv(directories, gene_names_of_interest)

# Print results
for gene, files in gene_files.items():
    print(f"\nCSV files containing the gene '{gene}':")
    for file in files:
        print(file)
CSV files containing the gene 'Notch1':
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Ovary_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Lung_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Spleen_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Liver_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Kidney_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/BM_unregulated_annotated_updated.csv

CSV files containing the gene 'Cebpb':
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/downregulated_genes_updated/BM_downregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Ovary_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Lung_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Spleen_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Liver_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Kidney_unregulated_annotated_updated.csv

CSV files containing the gene 'Junb':
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/upregulated_genes_updated/Liver_upregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/downregulated_genes_updated/BM_downregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Ovary_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Lung_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Spleen_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Kidney_unregulated_annotated_updated.csv

CSV files containing the gene 'Il1b':
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/upregulated_genes_updated/Liver_upregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/downregulated_genes_updated/BM_downregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Lung_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Spleen_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Kidney_unregulated_annotated_updated.csv

CSV files containing the gene 'Klf2':
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Ovary_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Lung_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Spleen_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Liver_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Kidney_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/BM_unregulated_annotated_updated.csv

CSV files containing the gene 'Zfp36':
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/downregulated_genes_updated/BM_downregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Ovary_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Lung_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Spleen_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Liver_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Kidney_unregulated_annotated_updated.csv

CSV files containing the gene 'Zfp36l1':
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Ovary_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Lung_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Spleen_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Liver_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Kidney_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/BM_unregulated_annotated_updated.csv

CSV files containing the gene 'Itgb2':
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/downregulated_genes_updated/BM_downregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Ovary_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Lung_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Spleen_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Liver_unregulated_annotated_updated.csv
/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated/Kidney_unregulated_annotated_updated.csv
In [45]:
import os
import pandas as pd

# Function to search for a list of gene names within CSV files in directories and classify them
def find_genes_in_csv(directories, gene_names):
    gene_files = {gene: {'Upregulated': [], 'Unregulated': [], 'Downregulated': []} for gene in gene_names}
    
    for directory in directories:
        category = 'Unregulated'  # Default category
        if 'upregulated' in directory.lower():
            category = 'Upregulated'
        elif 'downregulated' in directory.lower():
            category = 'Downregulated'
        
        for root, _, files in os.walk(directory):
            csv_files = [f for f in files if f.endswith('.csv')]
            for file in csv_files:
                file_path = os.path.join(root, file)
                try:
                    # Read the CSV file into a DataFrame
                    df = pd.read_csv(file_path)
                    
                    # Check if Gene.name column exists
                    if 'Gene.name' in df.columns:
                        for gene in gene_names:
                            if gene in df['Gene.name'].values:
                                # Extract tissue name from the file name
                                tissue = file.replace('_annotated_updated.csv', '').split('_')[0]
                                gene_files[gene][category].append(tissue)
                except Exception as e:
                    print(f"Error reading {file}: {e}")

    return gene_files

# List of directories to search
directories = [
    '/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/upregulated_genes_updated',
    '/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/downregulated_genes_updated',
    '/mnt/work_1/gsgeorge/mus_musculus/ARE_SCORE_ALL/unregulated_genes_updated'
]

# List of genes to search for
gene_names_of_interest = [
    'Notch1', 
    'Cebpb',  # CCAAT/enhancer binding protein-beta
    'Junb', 
    'Il1b', 
    'Klf2', 
    'Zfp36', 
    'Zfp36l1', 
    'Itgb2'
]

# Find CSV files containing the gene names
gene_files = find_genes_in_csv(directories, gene_names_of_interest)

# Print results
for gene, categories in gene_files.items():
    print(f"\n{gene}")
    for category, tissues in categories.items():
        print(f"{category}:")
        print(f"[{', '.join(tissues)}]" if tissues else "[None]")
Notch1
Upregulated:
[None]
Unregulated:
[Ovary, Lung, Spleen, Liver, Kidney, BM]
Downregulated:
[None]

Cebpb
Upregulated:
[None]
Unregulated:
[Ovary, Lung, Spleen, Liver, Kidney]
Downregulated:
[BM]

Junb
Upregulated:
[Liver]
Unregulated:
[Ovary, Lung, Spleen, Kidney]
Downregulated:
[BM]

Il1b
Upregulated:
[Liver]
Unregulated:
[Lung, Spleen, Kidney]
Downregulated:
[BM]

Klf2
Upregulated:
[None]
Unregulated:
[Ovary, Lung, Spleen, Liver, Kidney, BM]
Downregulated:
[None]

Zfp36
Upregulated:
[None]
Unregulated:
[Ovary, Lung, Spleen, Liver, Kidney]
Downregulated:
[BM]

Zfp36l1
Upregulated:
[None]
Unregulated:
[Ovary, Lung, Spleen, Liver, Kidney, BM]
Downregulated:
[None]

Itgb2
Upregulated:
[None]
Unregulated:
[Ovary, Lung, Spleen, Liver, Kidney]
Downregulated:
[BM]